Category Archives: Genetic Engineering

REHOVOT, Israel, June 4, 2021 /PRNewswire/ --CollPlant (NASDAQ: CLGN), a regenerative and aesthetic medicine company, announced today the uplisting of its ordinary shares to the Nasdaq Global Select Market effective at the open of market today, Friday, June 4, 2021.

CollPlant's ordinary shares now trade under the Company's current ticker symbol "CLGN" and the Company's American Depositary Shares (ADSs) have been mandatorily cancelled and exchanged for ordinary shares at a one-for-one ratio.Shareholders holding their ADSs in book-entry or through a bank, broker, or other nominee form do not need to take any action in connection with the mandatory exchange.

"We are pleased to complete this important milestone and believe that our current and future shareholders will benefit from our Nasdaq Global Marketstatus and the transition to ordinary shares," stated Yehiel Tal, CollPlant CEO.

A listing on the Nasdaq Global Marketis considered an indicator of status and success for companies that qualify for listing. Listed companies must satisfy stringent financial, liquidity and corporate governance requirements, both initially and on an ongoing basis.

About CollPlant

CollPlant is a regenerative and aesthetic medicine company focused on 3D bioprinting of tissues and organs, and medical aesthetics. The Company's products are based on its rhCollagen (recombinant human collagen) produced with CollPlant's proprietary plant based genetic engineering technology. These products address indications for the diverse fields of tissue repair, aesthetics, and organ manufacturing, and are ushering in a new era in regenerative and aesthetic medicine. CollPlant recently entered into a development and global commercialization agreement for dermal and soft tissue fillers with Allergan, an AbbVie company, the global leader in the dermal filler market.

For more information, visithttp://www.collplant.com.

Safe Harbor Statements

This press release may include forward-looking statements. Forward-looking statements may include, but are not limited to, statements relating to CollPlant's objectives plans and strategies, as well as statements, other than historical facts, that address activities, events or developments that CollPlant intends, expects, projects, believes or anticipates will or may occur in the future. These statements are often characterized by terminology such as "believes," "hopes," "may," "anticipates," "should," "intends," "plans," "will," "expects," "estimates," "projects," "positioned," "strategy" and similar expressions and are based on assumptions and assessments made in light of management's experience and perception of historical trends, current conditions, expected future developments and other factors believed to be appropriate. Forward-looking statements are not guarantees of future performance and are subject to risks and uncertainties that could cause actual results to differ materially from those expressed or implied in such statements. Many factors could cause CollPlant's actual activities or results to differ materially from the activities and results anticipated in forward-looking statements, including, but not limited to, the following: the Company's history of significant losses, its ability to continue as a going concern, and its need to raise additional capital and its inability to obtain additional capital on acceptable terms, or at all; the impact of the COVID-19 pandemic; the Company's expectations regarding the timing and cost of commencing clinical trials with respect to tissues and organs which are based on its rhCollagen based BioInk and products for medical aesthetics; the Company's ability to obtain favorable pre-clinical and clinical trial results; regulatory action with respect to rhCollagen based BioInk and medical aesthetics products including but not limited to acceptance of an application for marketing authorization review and approval of such application, and, if approved, the scope of the approved indication and labeling; commercial success and market acceptance of the Company's rhCollagen based products in 3D Bioprinting and medical aesthetics; the Company's ability to establish sales and marketing capabilities or enter into agreements with third parties and its reliance on third party distributors and resellers; the Company's ability to establish and maintain strategic partnerships and other corporate collaborations; the Company's reliance on third parties to conduct some or all aspects of its product manufacturing; the scope of protection the Company is able to establish and maintain for intellectual property rights and the Company's ability to operate its business without infringing the intellectual property rights of others; the overall global economic environment; the impact of competition and new technologies; general market, political, and economic conditions in the countries in which the Company operates; projected capital expenditures and liquidity; changes in the Company's strategy; and litigation and regulatory proceedings. More detailed information about the risks and uncertainties affecting CollPlant is contained under the heading "Risk Factors" included in CollPlant's most recent annual report on Form 20-F filed with the SEC, and in other filings that CollPlant has made and may make with the SEC in the future. The forward-looking statements contained in this press release are made as of the date of this press release and reflect CollPlant's current views with respect to future events, and CollPlant does not undertake and specifically disclaims any obligation to update or revise any forward-looking statements, whether as a result of new information, future events or otherwise.

Contact at CollPlant:Eran RotemDeputy CEO & Chief Financial OfficerTel: + 972-73-2325600Email:[emailprotected]

SOURCE CollPlant

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CollPlant Announces Effectiveness of Uplisting to the Nasdaq Global Market; Ordinary Shares Replace ADSs - PRNewswire

DetailsCategory: DNA RNA and CellsPublished on Sunday, 06 June 2021 12:38Hits: 418

8 of 11 Patients in Dose Escalation Cohorts 2 and 3 Achieved Objective Response

6 of 11 Patients Achieved Complete Response, including 2 Patients Previously Treated with Autologous CD19 CAR T-cell Therapy

Favorable FT516 Safety Profile Was Observed; No FT516-related Serious Adverse Events or FT516-related Grade 3 or Greater Adverse Events

Outpatient Treatment Regimen Was Well-tolerated; No Events of Any Grade of Cytokine Release Syndrome, Immune Effector Cell-Associated Neurotoxicity Syndrome, or Graft-vs-Host Disease

SAN DIEGO, CA, USA I June 04, 2021 I Fate Therapeutics, Inc. (NASDAQ: FATE), a clinical-stage biopharmaceutical company dedicated to the development of programmed cellular immunotherapies for cancer, today highlighted positive interim Phase 1 data from the Companys FT516 program for patients with relapsed / refractory B-cell lymphoma at the 2021 American Society of Clinical Oncology (ASCO) Annual Meeting being held virtually June 4-8, 2021. FT516 is the Companys universal, off-the-shelf natural killer (NK) cell product candidate derived from a clonal master induced pluripotent stem cell (iPSC) line engineered with a novel high-affinity, non-cleavable CD16 (hnCD16) Fc receptor, which is designed to maximize antibody-dependent cellular cytotoxicity (ADCC), a potent anti-tumor mechanism by which NK cells recognize, bind and kill antibody-coated cancer cells. The ongoing Phase 1 dose-escalation study of FT516 is currently enrolling patients in the fourth dose cohort of 900 million cells per dose.

As of the data cutoff date of March 11, 2021, four patients in the second dose cohort of 90 million cells per dose and seven patients in the third dose cohort of 300 million cells per dose were evaluable for assessment of safety and efficacy. Eight of eleven patients achieved an objective response, including six patients who achieved a complete response, as assessed by PET-CT scan per Lugano 2014 criteria (see Table 1). Patients had received a median of three prior lines of therapy and a median of two prior lines containing CD20-targeted therapy. Of the eleven patients, eight patients had aggressive B-cell lymphoma, five patients were refractory to their most recent prior therapy, and four patients were previously treated with autologous CD19 CAR-T cell therapy.

These additional data from our Phase 1 study of FT516 administered off-the-shelf in the outpatient setting continue to reinforce its differentiated safety profile and underscore its potential clinical benefit, said Wayne Chu, M.D., Senior Vice President of Clinical Development of Fate Therapeutics. Based on the favorable therapeutic profile of FT516 that continues to emerge and the potential to treat patients on-demand without delay, we plan to initiate multiple indication-specific, dose-expansion cohorts for patients with B-cell lymphomas to broadly assess FT516 in combination with CD20-targeted monoclonal antibody regimens, including those used as standard-of-care in earlier-line settings.

The ongoing Phase 1 clinical trial in relapsed / refractory B-cell lymphoma is assessing FT516 in an off-the-shelf treatment regimen of up to two cycles, with each cycle consisting of three days of conditioning chemotherapy (500 mg/m2 of cyclophosphamide and 30 mg/m2 of fludarabine), a single-dose of rituximab (375 mg/m2), and three weekly doses of FT516 each with IL-2 cytokine support. The FT516 treatment regimen is designed to be administered in the outpatient setting.

Safety DataNo dose-limiting toxicities, and no FT516-related serious adverse events or FT516-related Grade 3 or greater adverse events, were observed. The FT516 treatment regimen was well tolerated, and no treatment-emergent adverse events (TEAEs) of any grade of cytokine release syndrome, immune effector cell-associated neurotoxicity syndrome, or graft-versus-host disease were reported by investigators (see Table 2). All Grade 3 or greater TEAEs were consistent with lympho-conditioning chemotherapy and underlying disease. Of note, a Grade 3 or greater TEAE of infection was reported in one patient only. There were no discontinuations due to adverse events, and no patients withdrew from the study except in the setting of disease progression. In addition, no evidence of anti-product T- or B-cell mediated host-versus-product alloreactivity was detected, supporting the potential to safely administer up to six doses of FT516 in the outpatient setting without the need for patient matching.

Activity DataAs of the data cutoff date of March 11, 2021, eleven relapsed / refractory patients in the second and third dose cohorts were evaluable for assessment of safety and efficacy. Of the eleven patients, nine patients completed both FT516 treatment cycles and eight patients achieved an objective response, including six patients who achieved a complete response, as assessed by PET-CT scan per Lugano 2014 criteria. Notably, two of four patients previously treated with autologous CD19 CAR-T cell therapy achieved a complete response. Two patients showed progressive disease following the first FT516 treatment cycle and discontinued treatment. The Company previously reported that two patients treated in the first dose cohort (30 million cells per dose) showed progressive disease.

Patient Case StudyThe ASCO presentation featured a case study of a 36-year old male with triple-hit, high-grade B-cell lymphoma with rearrangements of MYC, BCL2, and BCL6 genes. The patient was refractory to all prior lines of therapy with the exception of autologous CD19 CAR T-cell therapy, for which a complete response of two months duration was achieved. The patient was most recently refractory to an investigational CD20-targeted T-cell engager and presented with bulky lymphadenopathy with the largest lesion measuring approximately 10 centimeters. The first FT516 treatment cycle resulted in a complete response with resolution of all metabolically active disease and 85% reduction in the size of target lesions. Subsequent to the data cutoff date of March 11, 2021, the patient completed a second FT516 treatment cycle after which the response assessment continued to show complete response.

As of March 11, 2021 database entry. Data subject to source document verification.CR = Complete Response; PR = Partial Response; PD = Progressive DiseaseCAR = Chimeric antigen receptor; DH/DE = Double-hit / double expressor; DLBCL = Diffuse large B-cell lymphoma; FL = Follicular lymphoma; Gr = Grade; HGBCL = High-grade B-cell lymphoma; iNHL = Indolent non-Hodgkin lymphoma; TH = Triple-hit; Transformed iNHL = Aggressive B-cell lymphoma transformed from indolent non-Hodgkin lymphoma1 Cycle 2 Day 29 protocol-defined response assessment per Lugano 2014 criteria2 Subject did not proceed to Cycle 23 Confirmed DLBCL (transformation from Gr3A FL) subsequent to the data cutoff date of March 11, 20214 Cycle 2 Day 29 protocol-defined response assessment reported subsequent to the data cutoff date of March 11, 2021

CRS = Cytokine Release Syndrome; DL = Dose Level; GvHD = Graft vs. Host Disease; ICANS = Immune Cell-Associated Neurotoxicity Syndrome;M = Million; SAE = Serious Adverse Event; TEAE = Treatment-Emergent Adverse Event1 Includes two subjects in the first dose cohort of 30 million cells per dose

About Fate Therapeutics iPSC Product PlatformThe Companys proprietary induced pluripotent stem cell (iPSC) product platform enables mass production of off-the-shelf, engineered, homogeneous cell products that are designed to be administered with multiple doses to deliver more effective pharmacologic activity, including in combination with other cancer treatments. Human iPSCs possess the unique dual properties of unlimited self-renewal and differentiation potential into all cell types of the body. The Companys first-of-kind approach involves engineering human iPSCs in a one-time genetic modification event and selecting a single engineered iPSC for maintenance as a clonal master iPSC line. Analogous to master cell lines used to manufacture biopharmaceutical drug products such as monoclonal antibodies, clonal master iPSC lines are a renewable source for manufacturing cell therapy products which are well-defined and uniform in composition, can be mass produced at significant scale in a cost-effective manner, and can be delivered off-the-shelf for patient treatment. As a result, the Companys platform is uniquely designed to overcome numerous limitations associated with the production of cell therapies using patient- or donor-sourced cells, which is logistically complex and expensive and is subject to batch-to-batch and cell-to-cell variability that can affect clinical safety and efficacy. Fate Therapeutics iPSC product platform is supported by an intellectual property portfolio of over 350 issued patents and 150 pending patent applications.

About FT516FT516 is an investigational, universal, off-the-shelf natural killer (NK) cell cancer immunotherapy derived from a clonal master induced pluripotent stem cell (iPSC) line engineered to express a novel high-affinity 158V, non-cleavable CD16 (hnCD16) Fc receptor, which has been modified to prevent its down-regulation and to enhance its binding to tumor-targeting antibodies. CD16 mediates antibody-dependent cellular cytotoxicity (ADCC), a potent anti-tumor mechanism by which NK cells recognize, bind and kill antibody-coated cancer cells. ADCC is dependent on NK cells maintaining stable and effective expression of CD16, which has been shown to undergo considerable down-regulation in cancer patients. In addition, CD16 occurs in two variants, 158V or 158F, that elicit high or low binding affinity, respectively, to the Fc domain of IgG1 antibodies. Scientists from the Company have shown in a peer-reviewed publication (Blood. 2020;135(6):399-410) that hnCD16 iPSC-derived NK cells, compared to peripheral blood NK cells, elicit a more durable anti-tumor response and extend survival in combination with anti-CD20 monoclonal antibodies in an in vivo xenograft mouse model of human lymphoma. Numerous clinical studies with FDA-approved tumor-targeting antibodies, including rituximab, trastuzumab and cetuximab, have demonstrated that patients homozygous for the 158V variant, which is present in only about 15% of patients, have improved clinical outcomes. FT516 is being investigated in a multi-dose Phase 1 clinical trial as a monotherapy for the treatment of acute myeloid leukemia and in combination with CD20-targeted monoclonal antibodies for the treatment of advanced B-cell lymphoma (NCT04023071). Additionally, FT516 is being investigated in a multi-dose Phase 1 clinical trial in combination with avelumab for the treatment of advanced solid tumor resistant to anti-PDL1 checkpoint inhibitor therapy (NCT04551885).

About Fate Therapeutics, Inc.Fate Therapeutics is a clinical-stage biopharmaceutical company dedicated to the development of first-in-class cellular immunotherapies for patients with cancer. The Company has established a leadership position in the clinical development and manufacture of universal, off-the-shelf cell products using its proprietary induced pluripotent stem cell (iPSC) product platform. The Companys immuno-oncology pipeline includes off-the-shelf, iPSC-derived natural killer (NK) cell and T-cell product candidates, which are designed to synergize with well-established cancer therapies, including immune checkpoint inhibitors and monoclonal antibodies, and to target tumor-associated antigens using chimeric antigen receptors (CARs). Fate Therapeutics is headquartered in San Diego, CA. For more information, please visit http://www.fatetherapeutics.com.

SOURCE: Fate Therapeutics

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Fate Therapeutics Highlights Positive Interim Data from its Phase 1 Study of FT516 in Combination with Rituximab for B-cell Lymphoma at 2021 ASCO...

Global Genome Editing/Genome Engineering Market is valued approximately USD 4.4 billion in 2019 and is anticipated to grow with a healthy growth rate of more than 17.00 % over the forecast period 2020-2027.

Genome Engineering technique is used for deletion, insertion and modification of genome of a microorganism. Genome editing/genome engineering plays an integral role in modern-day biology and is widely used in the biopharmaceutical and biotechnology industry to alter the genome of microorganism to perform process such as fermentation which yield desired products. Also, this approach is majorly used for understanding DNA in cells of organism to have a better understanding of their biology, to treat infectious and autoimmune diseases. Availability of government funding and growth in the number of genomics projects are the few factors responsible for growth of the market over the forecast period of 2020-2027. For Instance: in 2017, in Canada, as per the University of Guelph, University of Genome Canada Bioinformatics and Computational Biology along with other eligible sources provide funding of $12 million for different genomics-based research projects. Similarly, according the Japan Agency for Medical Research and Development Organization, in 2017, government of Japan has introduced various initiatives such as Tohoku Medical Megabank project, Platform Program for promotion of Genome Medicine etc., these projects are inclined to provide research infrastructure.

Request for a FREE sample of this market research report@ https://www.reportocean.com/industry-verticals/sample-request?report_id=bw1334

Such government initiatives would increase the need for genome editing and genome engineering. Also, the key players of global Genome Editing/Genome Engineering market have adopted various strategies to gain competitive advantage including product launch, innovation, technological advancements, investment, funding and others. However, high equipment cost is the major factor restraining the growth of global Genome Editing/Genome Engineering market during the forecast period.

The regional analysis of global Genome Editing/Genome Engineering market is considered for the key regions such as Asia Pacific, North America, Europe, Latin America and Rest of the World. North America is the leading/significant region across the world in terms of market share as the region is one of the most significant markets for development of gene therapy in the US, the the rising prevalence of infectious diseases and cancer, increasing use of genetically modified crops, and the availability of research grants and funding. Whereas Asia-Pacific is also anticipated to exhibit highest growth rate / CAGR over the forecast period 2020-2027.

Major market player included in this report are:Thermo Fisher ScientificMerckHorizon DiscoveryGenscriptSangamo TherapeuticsLonzaEditas MedicineCrispr TherapeuticsEurofins ScientificPrecision Biosciences

The objective of the study is to define market sizes of different segments & countries in recent years and to forecast the values to the coming eight years. The report is designed to incorporate both qualitative and quantitative aspects of the industry within each of the regions and countries involved in the study. Furthermore, the report also caters the detailed information about the crucial aspects such as driving factors & challenges which will define the future growth of the market. Additionally, the report shall also incorporate available opportunities in micro markets for stakeholders to invest along with the detailed analysis of competitive landscape and product offerings of key players. The detailed segments and sub-segment of the market are explained below:

By Technology:CRISPRTALENZFNANTISENSEOther technologiesBy Product and Services:Reagents and ConsumablesSoftware & systemsServicesBy Application:Cell line EngineeringGenetic EngineeringDiagnostics ApplicationsDrug discovery and developmentOthersBy End-user:Pharmaceuticals CompaniesBiotechnology CompaniesAcademic and Government research institutesBy Region:North AmericaU.S.CanadaEuropeUKGermanyFranceSpainItalyROE

Asia PacificChinaIndiaJapanAustraliaSouth KoreaRoAPACLatin AmericaBrazilMexicoRest of the World

Furthermore, years considered for the study are as follows:

Historical year 2017, 2018Base year 2019Forecast period 2020 to 2027

Send a request to Report Ocean to understand the structure of the complete report @https://www.reportocean.com/industry-verticals/sample-request?report_id=bw1334

Target Audience of the Global Genome Editing/Genome Engineering Market in Market Study:

Key Consulting Companies & AdvisorsLarge, medium-sized, and small enterprisesVenture capitalistsValue-Added Resellers (VARs)Third-party knowledge providersInvestment bankersInvestors

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Genome Editing/Genome Engineering Market expectation surges with rising demand and changing trends by industry analysis through 2027 The Manomet...

The World Health Organisation maintains the coronavirus most likely arose in bats, and then spread to humans via an as-yet unidentified intermediary animal. Image / CDC

Despite the "lab leak theory" taking flight again in the US media, global scientists still point out there's little evidence to suggest the SARS-CoV-2 virus originated in a Wuhan laboratory and a large amount to suggest it came from nature. Otago University virologist Dr Jemma Geoghegan and Massey University infectious disease ecologist Professor David Hayman set out three reasons why a lab-made pandemic remains extremely unlikely.

There's a strong precedent for coronaviruses that have become "zoonotic", or jumped from animals to humans.

There have been seven that scientists are aware of including SARS-CoV and MERS-CoV and bats are thought to have been involved in most of these.

"And five of these human coronaviruses have emerged in the last 20 years," Geoghegan said.

While the source of the SARS-CoV-2 virus still hasn't been pinned down, she said that wasn't unusual.

"In fact, we don't know where most of the viruses that infect us have come from," she said.

"This is why we need to sample more viruses in nature and expand our knowledge of the diversity of viruses that exist."

Hayman added that, now that scientists were looking even harder, more cases of infections from newly-detected or "novel" coronaviruses were coming to light.

"One was identified in pneumonia patients in Malaysia where people were living in villages in close contact with domestic and wild animals."

While the pandemic found the world poorly prepared for it, scientists had been warning for years that the growing interaction between animals and nature particularly through habitat destruction had been raising the risk of a catastrophe like Covid-19.

Just like the first Sars coronavirus, the first cases of Sars-Cov-2 were associated with an animal market this time in Wuhan, China.

The World Health Organisation's report identified that live animals like ferret-badgers and rabbits were being traded in these markets.

"These animals could provide an intermediate host for the virus to jump to humans," Geoghegan said.

"It's exactly the type of place you'd expect a zoonosis event to happen."

Hayman added that some of the farmed species had complex commodity chains.

That meant the farms could well be in places where there was a greater diversity of bat viruses than in Wuhan, where the pandemic appeared to have started.

"And we have seen how SARS-CoV-2 can be maintained in farmed fur animals, such as the very large outbreaks in farmed mink in Europe, which led to substantial mink to mink transmission as well as mink-to-human transmission."

Other evidence also shows that this type of coronavirus has existed in bats for decades and the SARS-CoV-2 genome sequence happened to be 96 per cent identical to a coronavirus found in horseshoe bats.

In one of the earliest major studies into the virus, scientists analysed its genetic template for spike proteins or armatures on the outside of the virus that it used to grab and penetrate the outer walls of human and animal cells.

More specifically, they focused on two important features of the spike protein.

Those were its receptor-binding domain (RBD) - a kind of grappling hook that grips on to host cells - and what's called the cleavage site, a molecular can opener that allows the virus to crack open and enter host cells.

They found the RBD portion of the SARS-CoV-2 spike proteins had evolved to effectively target a molecular feature on the outside of human cells called ACE2 - a receptor involved in regulating blood pressure.

The SARS-CoV-2 spike protein was so effective at binding the human cells, in fact, that the scientists concluded it was the result of natural selection, and not the product of genetic engineering that some theorists have suspected.

The idea of natural evolution was given further credence by data on the virus' backbone - its overall molecular structure.

If someone were seeking to engineer a new coronavirus as a pathogen, they would have constructed it from the backbone of a virus known to cause illness.

But the scientists found that the backbone differed greatly from those of already known coronaviruses.

It turned out to mostly resemble related viruses found in bats and pangolins - scaly-skinned mammals that are prized delicacies in China.

That led scientists to suspect one of two possible scenarios.

In one scenario, the virus evolved to its current state through natural selection in an animal host and then jumped to humans.

Yet there were no documented cases of direct bat-human transmission, suggesting that an intermediate host was likely involved between bats and humans.

In this scenario, both of the distinctive features of SARS-CoV-2's spike protein - the RBD portion that binds to cells and the cleavage site that opens the virus up - would have evolved to their current state before entering humans.

In this case, the current epidemic would probably have emerged rapidly as soon as humans were infected, as the virus would have already evolved the features that made it pathogenic, or able to spread between people.

In the other proposed scenario, a non-pathogenic version of the virus jumped from an animal host into humans and then evolved to its current state within the human population.

A coronavirus from a pangolin could possibly have been transmitted to a human, either directly or through an intermediary host such as civets or ferrets.

After that, the other distinct spike protein characteristic of SARS-CoV-2 - the cleavage site - could have evolved within a human host, or possibly among a group of people, before the outbreak kicked off.

Geoghegan said the idea that the cleavage site was so unusual that it must have been engineered was "totally false".

She said that assumed an amino acid sequence within the site called PRRAR had been created in a lab.

Yet these cleavage sites had been found in other coronaviruses - even with the exact same "PRRAR" insert.

"It's a totally bonkers argument," Hayman added.

"Similarly, people really need to understand that these viruses do recombine. For example, the novel virus from Malaysia that was recently detected seems to be a recombinant of a cat and dog viruses, which were also previously not known."

The slightly more plausible alternative lab leak theory was that scientists could simply have been growing a culture of the virus, and it escaped from there.

But that would've had to assume the virus could have leaked from a secure research facility and also neglected the fact that the virus' feature of entry and infection markedly diminished in a lab culture setting.

And then, as Dr Jonathan Stoye, group leader of the Retrovirus-Host Interactions Laboratory at the UK's Francis Crick Institute, pointed out, the virus' spread around the world didn't gel with the lab-grown theory.

"The genome of SARS-CoV-2 shows more than 1000 individual differences from its closest known relative," he said.

"Given the rate of nucleotide change observed in virus spreading through the human population over the past year it seems extremely improbable, perhaps impossible, that changes spanning such an evolutionary distance could have occurred during virus growth in a lab.

"It therefore remains most likely that the immediate ancestor to SARS-CoV-2 exists in the wild and is still to be found."

Scientists aren't saying the possibility of a lab leak should be entirely ruled out on the contrary, many argue that it should be comprehensively investigated.

But more than a year after the outbreak, the weight of evidence overwhelmingly points to a natural source, while there's little to suggest the virus came from a lab.

Part of the lab leak theory is predicated on the fact that the Wuhan Institute of Virology has carried out extensive work on coronaviruses in bats.

While the institute didn't shared its lab records with a team of WHO investigators, there's as yet been no evidence that any samples of the virus were kept there before it was first reported, nor were there any viruses that could have combined to create it.

Much of the recent coverage has been fueled by a US intelligence report that stated several researchers had become sick with "symptoms consistent with both Covid-19 and common seasonal illness".

A top director at the institute has rejected this inference, reporting that all staff have tested negative for Covid-19 antibodies, and that there'd been no turnover of staff in the coronavirus team.

"Why would these scientists be working on a random secret bat virus and not have published anything on it previously?" Geoghegan said.

"Those that do gain-of-function experiments work on really well characterised viruses, and there would be really close genetic relatives of the virus published before.

"But for SARS-CoV-2, there isn't. Even the closest viruses in bats and pangolins are too divergent to be a starting point."

Hayman added that every expert in the area would have asked themselves, "what if it was that lab?"

"We ask ourselves, 'What would we see, what evidence would we need to support it?', and even, 'have we been lied to?'. We have, most of us, agonised over the possibilities.

"But right now the only way that this can be true is if there is a massive conspiracy, because while the lab pathway is a potential pathway, nothing published or reliably reported by China to date supports a lab escape at all, and there is a huge amount of data to support this being a natural event.

"This discussion would have gone away if it hadn't have happened in China. But unfortunately finding conclusive evidence of either is very unlikely."

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Covid 19 coronavirus: Why the lab leak theory is still unlikely - New Zealand Herald

Markeshaw Tiruneh G/Medhin,1 Endeshaw Chekol Abebe,2 Tekeba Sisay,3 Nega Berhane,3 Tesfahun Bekele Snr,1 Tadesse Asmamaw Dejenie1

1Department of Biochemistry, School of Medicine, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 2Department of Biochemistry, College of Health Sciences, Debre Tabor University, Debre Tabor, Ethiopia; 3Institute of Biotechnology, College of Natural Science, University of Gondar, Gondar, Ethiopia

Correspondence: Markeshaw Tiruneh G/Medhin Tel +251922712112Email [emailprotected]

Abstract: Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins are referred to as CRISPR-Cas9. Bacteria and archaea have an adaptive (acquired) immune system. As a result, developing the best single regulated RNA and Cas9 endonuclease proteins and implementing the method in clinical practice would aid in the treatment of diseases of various origins, including lung cancers. This seminar aims to provide an overview of CRISPR-Cas9 technology, as well as current and potential applications and perspectives for the method, as well as its mechanism of action in lung cancer therapy. This technology can be used to treat lung cancer in two different ways. The first approach involves creating single directed RNA and Cas9 proteins and then distributing them to cancer cells using suitable methods. Single directed RNA looks directly at the lungs mutated epidermal growth factor receptor and makes a complementary match, which is then cleaved with Cas9 protein, slowing cancer progression. The second method is to manipulate the expression of ligand-receptors on immune lymphocytic cells. For example, if the CRISPR-Cas9 system disables the expression of cancer receptors on lymphocytes, it decreases the contact between the tumor cell and its ligand-receptor, thus slowing cancer progression.

Keywords: CRISPR, Cas9, CRISPR-Cas9 technology, cancer, lung cancer, cancer treatment

The word CRISPR-Cas9 refers to Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins.15 CRISPR-Cas9 system is a kind of acquired immunity possessed by most bacteria and archaea (prokaryotes) to act against their enemies (bacteriophages).4,6 It is a ribonucleic acid (RNA) guided, convenient, and versatile endonuclease platform for site-specific genome editing,1,7,8 which can play a tremendous role in the application of cancer therapy.1 The application of this technology can be used to resolve mutations and to introduce site-specific therapeutic genes in human cells so that, correcting disease-causing mutations, and alleviate disease-related symptoms. This system is also a useful tool for delineating molecular mechanisms involving hematological malignancies.4 Sequence-specific gene editing using CRISPR-Cas9 shows promise as a novel therapeutic approach for the treatment of a variety of diseases that currently lack effective treatments like cancers.3,9 To accomplish its task, it requires Cas9 DNA endonuclease protein and single guided RNA (sgRNA) that can produce precise gene matching for editing and correction techniques.2 So the system has enabled easy manipulation of genes for the scientific community by making the hybrid to the target sequence and cleaving the double-strand DNA.10

Additionally, the CRISPR-Cas9 technology is increasingly feasible to overcome drug resistance in breast cancer therapy and will become an essential tool for personalized medicine.4 It is a technological breakthrough that facilitates the ability to change nucleic acids,11 and with continued improvement in the function, the system can help to develop best treatment options to a variety of genetic disease which affects several tissues in our body.12 Gene manipulation using CRISPR-Cas9 system has revolutionized and made it easy to study the work of genes and importantly opens the new era of treatment mechanisms for different disease conditions including cancer.13 Technologies like this are a simple and efficient method of targeting the required DNA regions.14 Thus, scientists have designed two main components of the system for easy detection and alteration of gene function one component is a protein Cas9 that enzymatically cleave the desired gene and the sgRNA which scans and determines where the gene of interest will be cleaved by Cas9 protein.3,12,15 The system has been scientifically optimized and developed to regulate expression of the gene, modify and edit the desired locus and this makes the technology of choice seen by the scientific community to treat or edit disease-causing mutations more efficiently than ever before. Furthermore, its application is encouraging for more vigorous gene therapy in clinical setups.16 Based on the discovery, there are three main types of CRISPR-Cas9 system (I to III) and with three additional types (IV to VI) being identified more recently.17 They are different during the processes of immunity, adaptation, expression, and interference, each type acts in distinct mechanisms to ensure genetic manipulation. Type I employs a large complex of Cas9 proteins with distinct helicase and DNase activities, while type III employs repeat-associated mysterious proteins, which form a large Cas9 superfamily. Another classification is based on subunit effectors, with multi-subunit effector complexes being the most common. Types I, III, and IV are grouped in class 1. Those systems, on the other hand, that have a single subunit effector are categorized as class 2 comprising types II, V, and VI.17,18 Type II uses only a Single protein (Cas9) for its nuclease activity and has got more attention and adopted for genome engineering.5,17,19 Thus, the objective of this seminar is to introduce CRISPR-Cas9 technology and describe current applications and future perspectives of the system with its mechanism of action on lung cancer therapy.

The tracrRNA gene will be transcribed to tracrRNA, the crRNA gene will be transcribed to pre crRNA, and the Cas9 gene will be transcribed to Cas9 messenger RNA and converted to Cas9 protein, all of which will be post-transcribed and chopped off to form the mature CRISPR-Cas9 complex.20 Cas1 and Cas2 integrase, which are present in all CRISPR forms, catalyze spacer integration on the CRISPR array especially on the leader end of the repeat there will be a nucleophilic attack of the 3 OH of the protospacers followed by the same practice on the spacer end of the repeat.21

The CRISPR-Cas9 method has a variety of formulation methods for genome editing. The use of a plasmid-based CRISPR-Cas9 system encoding both the Cas9 protein and sgRNA from the same vector, which is necessary to avoid multiple transfections of different components of the technology, is the leading and possibly the easiest technique. The Cas9 protein and sgRNA will be expressed in the vector, which will form the sgRNA-Cas9 complex within cells to edit the target genomic sequences.3,12,15,18 The second approach involves combining Cas9 mRNA and sgRNA. The sgRNA-Cas9 complex will be formed when Cas9 mRNA is converted into Cas9 protein in cells. The third strategy is to deliver the in vitro assembled sgRNA-Cas9 complex directly to the cell.18

It is difficult to transmit nucleic acid in general, and CRISPR-Cas9 in particular, to the target tissue or cell. Physical and vector (viral or non-viral) approaches are two of the most widely used distribution strategies.11,22,23 Electroporation and microinjections are used in physical methods, while viral delivery strategies such as adeno associated virus (AAV) are widely used in vector-based methods since they are not disease-causing agents and can infect both dividing and non-dividing cells25 and lentivirus with inactivated integrase enzymes are under investigation.24

Another technique is lipofection (lipid-mediated nanoparticle transfection), which is possibly the most efficient CRISPR-Cas9 in vivo delivery method.22 This technique was further developed by26 and is currently being tested in clinical trials.13,24

The CRISPR-Cas9 system, as discussed, a little earlier, is made up of two main components that work together to accomplish its goal.19 The sgRNA contains crRNA, which scans and identifies the target DNA sequences that must be cleaved and corrected, and transactivated crRNA (tracrRNA), which recruits component two, the Cas9 protein DNA endonuclease, which can sense, identify, and establish site-specific double-strand DNA breaks (DSB).15 Because of its simplicity and convenience, the bacterial type II CRISPR-Cas9 system has been used for RNA-guided engineering nucleases.4,18 However, the proto-spacer adjacent motifs (PAM) sequences are required by the method. After recognition, two Cas9 domains cleave double-stranded DNA: the endonuclease domain named for characteristic histidine and asparagine residues (HNH) domain, which cleaves the complementary strand, and the endonuclease domain named for an E. coli protein involved in DNA repair (RuvC-like) domain, which cleaves the non-complementary strand.17 As a result, the host DNA repair machinery introduces numerous mutations such as substitutions, deletions, and insertions in the target genome, including non-homologous end joining (NHEJ) or homologous-dependent repair (HDR).1518 Another paper, CRISPR-Cas9 for Cancer Therapy: Hopes and Challenges, supports this theory by demonstrating that the sgRNA-Cas9 complex scans and anneals to the genomic target sequence with base-pairing complementarity and precisely cleaves double-stranded DNA of the target cell after identification of the protospacer adjacent motif (PAM) sequence adjacent to the target sequence. NHEJ or HDR pathways are activated as a result of double-strand breaks. NHEJ is an error-prone repair mechanism that results in indels (insertions or deletions) of random base pairs disrupting the target sequence in the absence of a homologous repair prototype with more specific repair mechanisms.23,27

Lung cancer is the major cause of death in the United States and a significant public health concern worldwide.5 In both developed and developing countries, it is a common cause of morbidity and mortality.28 According to a study conducted by the American Lung Cancer Society in 2015, lung cancer claims the lives of almost 150,000 people each year. However, surgery and radiation were used as treatment options. The treatment was later changed to selective Tyrosine kinase inhibitors (TKIs) like gefitinib and erlotinib to inhibit the tyrosine kinase activity of epidermal growth factor receptor, which has technical difficulties and nonspecific cytotoxicity (EGFR).29,30 Extracellular ligand binding, transmembrane, and intracellular tyrosine kinase domains are found in this membrane glycoprotein. When the ligand activator binds to the extracellular ligand domain, it transduces and initiates intracellular kinase activities, which cause cellular proliferation, neovascularization, invasion, and metastasis, as well as reduced apoptosis and glycolysis activation. These medications, however, have encountered drug resistance.28,29

The CRISPR-Cas9 device is the start of a new biotechnological era and a groundbreaking technology that is being used to treat lung cancer.6,29 The system works in two ways. The first is by designing sgRNA that looks for the mutated EGFR sequence, which is then accompanied by Cas9 protein. To do so, scientists created a CRISPR device that has complementary sequences with the mutated EGFR and introduced it into the patient, as mentioned earlier which has complementary sequences with the mutated EGFR and introducing this into the patient. As this complementary sequence binds to the mutated EGFR, the Cas9 protein (endonuclease) creates a double-stranded or single-stranded DNA break, depending on the type of enzyme used, followed by DNA repair mechanisms such as homologous or non-homologous DNA repair.29 If the receptor mutation is limited, there will be no contact between the ligand activator, resulting in no cell proliferation, neovascularization, or cancer metastasis, and the problem will be solved. The inhibition of EGFR by CRISPR-Cas9 increases the expression of major histocompatibility complex class I, which improves cytotoxic lymphocyte recognition and lysis of tumor cells.30,31 Off-target effects, which can induce genome instability, gene functional disturbances, and epigenetic alterations, are a challenge. Off-target effects of CRISPR-Cas9 systems, particularly when used for therapeutic purposes, should be minimized and precisely profiled. Off-target effects are separated into two categories: off-target binding and off-target cleavage. Cas9 can bind to target sequences that are partially complementary to sgRNA and inhibit target gene transcription without cleaving them.8 Off-target binding effects may thus be removed in traditional off-target identification approaches, such as using in vitro assembled sgRNA with a long-lasting association with cas9, which also has a high proportion of on-target and high efficiency for genome editing. Another technique is to use a Cas9 variant or modified Cas9 that can generate a single nick at one strand.23 So that the off-target effect is reduced.

The second, and equally significant, strategy for using this biotechnological method to treat lung cancer is to search for immune cells like lymphocytes. T cells are immune cells that are extracted from the blood of patients engaged in a clinical trial for lung cancer treatment in China, and then CRISPR-Cas9 is used to knock out a gene in the cells that encodes a protein called PD-1. The edited gene cells would then be propagated in the lab before being injected back into the patients bloodstream.6,25 Scientists took blood from the patient and extracted lymphocytes, which were then treated with a CRISPR-Cas9 gene-editing system containing a sgRNA sequence with a pattern identical to lymphocytes programmable death 1 protein (PD 1). When the system detects its target sequence, cas9 would sever the DNA, which is then repaired by cell repair mechanisms. When the expression of the PD 1 gene is blocked or disabled, cancerous cells lack the receptor on immune lymphocytes.6,25 As a result, if lymphocytes do not express the PD 1 receptor well, there will be less contact between the cancerous ligand and receptor, causing the T cell receptor to identify the problematic cell and perform its function. Naturally, these manipulated lymphocytes were screened for viability and lympho-proliferation to rule out new mutations, and only those cells that passed the test were returned to the patient.6,25 Furthermore, knocking out the PD-1 protein on immune cells is necessary for caspase activation, which is needed for programmed cell death and enhanced apoptosis in cancerous cells.31 It also concludes that PD-1-deficient cells have potent antitumor activity of cytotoxic lymphocytes. The hyperactivity of the manipulated T cells is one of the technologys drawbacks for use in this way6 and obtaining a safe and efficient delivery method, as well as some side effects Patients with advanced NSCLC with positive PD-1 expression were assigned to a Phase I clinical trial to assess the safety of CRISPR-Cas9-mediated knockout of PD-1 gene therapy in patients with metastatic non-small cell lung cancer. Nine patients were enrolled, and eight patients received PD-1 deficient T cell therapy, and the patients were manifested with PD-1 deficient T cell therapy.25 Patients undergoing PD-1 deficient T cell therapy, on the other hand, appeared to be healthy, and researchers recommended that broader studies be conducted to determine the most appropriate dosage and immune response.

In cancer biology, the CRISPR-Cas9 device has a bright future ahead of it,9, because it is a technology that is adaptable, simple, convenient and efficient.32,33 The method introduces a novel approach to cancer treatment by allowing for modifications to the genome of target cells, which was previously difficult to achieve.3436 the technologys versatility, effectiveness, and flexibility would make it the best form of cancer care in the future.4,37,38 It will affect cancer biology as a whole in the future,34 and if researchers have devised well-organized strategies and instruments for delivering the technology to the target cell or tissue, as well as effective methods and instructions for controlling and eliminating the technologys off-target effects.

The CRISPR-Cas9 device is a recent biotechnological breakthrough and scientific achievement. This technology has created a new treatment option for diseases of various origins, such as cancer and infectious disease. To solve the problem, the best sgRNA must be designed using a CRISPR tool (http://crispr.mit.edu) and its associated endonuclease cas9 protein against the target sequence. However, ethical concerns, the need for the best delivery strategies, and the risk of off-target effects are only a few of the problems that must be addressed. Since the technology is still in its infancy, researchers must devise simple methods and mechanisms to track and test its protection and efficacy. For a simple comparison, the benefits of this technology are simple, fast, relatively effective, relatively precise, and versatile, while the drawbacks are distribution is difficult, ethical problems are highly conservative, some off-target effects, and some adverse effects.

ATP, Adenosine triphosphate; CRISPR, Clustered regularly interspaced short palindromic repeat; CRISPR-Cas, Clustered regularly interspaced short palindromic repeat-associated; CrRNA, Clustered regularly interspaced short palindromic repeat ribonucleic acid; DNA, Deoxyribonucleic acid; DSB, Double-stranded break; EGFR, epidermal growth factor receptor; HDR, Homologous directed repair; mRNA, Messenger ribonucleic acid; NHEJ, Non-homologous end-joining; PD 1, Programmable death protein 1; RNA, Ribonucleic acid; SgRNA, Single guided ribonucleic acid; TracrRNA, Trans activating clustered regularly short palindromic repeat ribonucleic acid; TKIs, Tyrosine kinase inhibitors.

All authors made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work.

There is no funding to report.

The authors declare that they have no conflicts of interest for this work.

1. Liu T, Shen JK, Li Z, Choy E, Hornicek FJ, Duan Z. Development and potential applications of CRISPR-Cas9 genome editing technology in sarcoma. Cancer Lett. 2016;373(1):109118. doi:10.1016/j.canlet.2016.01.030

2. Yin H, Song CQ, Dorkin JR, et al. Therapeutic genome editing by combined viral and non-viral delivery of CRISPR system components in vivo. Nat Biotechnol. 2016;34(3):328333. doi:10.1038/nbt.3471

3. Zhen S, Li X. Oncogenic human papillomavirus: application of CRISPR/Cas9 therapeutic strategies for cervical cancer. Cell Physiol Biochem. 2017;44(6):24552466. doi:10.1159/000486168

4. Chen Y, Zhang Y. Application of the CRISPR/Cas9 system to drug resistance in breast cancer. Adv Sci. 2018;5(6):1700964. doi:10.1002/advs.201700964

5. Shen Q, Li J, Mai J, et al. Sensitizing non-small cell lung cancer to BCL-xL-targeted apoptosis. Cell Death Dis. 2018;9(10):13. doi:10.1038/s41419-018-1040-9

6. Castillo A. Gene editing for the treatment of lung cancer (CRISPR-Cas9). Colomb Med. 2016;47(4):178180. doi:10.25100/cm.v47i4.2856

7. Platt RJ, Chen S, Zhou Y, et al. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell. 2014;159(2):440455. doi:10.1016/j.cell.2014.09.014

8. Wen WS, Yuan ZM, Ma SJ, Xu J, Yuan DT. CRISPRCas9 systems: versatile cancer modeling platforms and promising therapeutic strategies. Int J Cancer. 2016;138(6):13281336. doi:10.1002/ijc.29626

9. Bhattacharjee R, Purkayastha KD, Adapa D, Choudhury A. CRISPR/Cas9 genome editing system in the diagnosis and treatment of cancer. J RNAi Gene Silencing. 2017;13:585591.

10. Gwiazda KS, Grier AE, Sahni J, et al. High-efficiency CRISPR/Cas9-mediated gene editing in primary human T-cells using mutant adenoviral E4orf6/E1b55k helper proteins. Mol Ther. 2016;24(9):15701580. doi:10.1038/mt.2016.105

11. Wang T, Wei JJ, Sabatini DM, Lander ES. Genetic screens in human cells using the CRISPR-Cas9 system. Science. 2014;343(6166):8084. doi:10.1126/science.1246981

12. Ran FA, Cong L, Yan WX, et al. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015;520(7546):186191. doi:10.1038/nature14299

13. Gori JL, Hsu PD, Maeder ML, Shen S, Welstead GG, Bumcrot D. Delivery and specificity of CRISPR/Cas9 genome editing technologies for human gene therapy. Hum Gene Ther. 2015;26(7):443451. doi:10.1089/hum.2015.074

14. Shinmyo Y, Tanaka S, Tsunoda S, Hosomichi K, Tajima A, Kawasaki H. CRISPR/Cas9-mediated gene knockout in the mouse brain using in utero electroporation. Sci Rep. 2016;6(1):13. doi:10.1038/srep20611

15. Ratan ZA, Son YJ, Haidere MF, et al. CRISPR-Cas9: a promising genetic engineering approach in cancer research. Ther Adv Med Oncol. 2018;10:1758834018755089. doi:10.1177/1758834018755089

16. Jamal M, Ullah A, Ahsan M, et al. Treating genetic disorders using state-of-the-art technology. Curr Issues Mol Biol. 2017;26:3346. doi:10.21775/cimb.026.033

17. Kim EJ, Kang KH, Ju JH. CRISPR-Cas9: a promising tool for gene editing on induced pluripotent stem cells. Korean J Intern Med. 2017;32(1):42. doi:10.3904/kjim.2016.198

18. Liu C, Zhang L, Liu H, Cheng K. Delivery strategies of the CRISPR-Cas9 gene-editing system for therapeutic applications. J Control Release. 2017;266:1726. doi:10.1016/j.jconrel.2017.09.012

19. Snchez-Rivera FJ, Jacks T. Applications of the CRISPRCas9 system in cancer biology. Nat Rev Cancer. 2015;15(7):387393. doi:10.1038/nrc3950

20. Lino CA, Harper JC, Carney JP, Timlin JA. Delivering CRISPR: a review of the challenges and approaches. Drug Deliv. 2018;25(1):12341257. doi:10.1080/10717544.2018.1474964

21. McGinn J, Marraffini LA. Molecular mechanisms of CRISPRCas spacer acquisition. Nat Rev Microbiol. 2019;17(1):712. doi:10.1038/s41579-018-0071-7

22. Finn JD, Smith AR, Patel MC, et al. A single administration of CRISPR/Cas9 lipid nanoparticles achieves robust and persistent in vivo genome editing. Cell Rep. 2018;22(9):22272235. doi:10.1016/j.celrep.2018.02.014

23. Martinez-Lage M, Puig-Serra P, Menendez P, Torres-Ruiz R, Rodriguez-Perales S. CRISPR/Cas9 for cancer therapy: hopes and challenges. Biomedicines. 2018;6(4):105. doi:10.3390/biomedicines6040105

24. Kolli N, Lu M, Maiti P, Rossignol J, Dunbar GL. Application of the gene-editing tool, CRISPR-Cas9, for treating neurodegenerative diseases. Neurochem Int. 2018;112:187196. doi:10.1016/j.neuint.2017.07.007

25. Lu Y. PD-1 knockout engineered T cells for metastatic non-small cell lung cancer. ClinicalTrials.gov. 2019. doi:10.1016/j.compcom.2006.05.002

26. Doudna JA. The promise and challenge of therapeutic genome editing. Nature. 2020;578(7794):229236. doi:10.1038/s41586-020-1978-5

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Read more from the original source:
Crispr-cas9 for the treatment of lung cancer | BTT - Dove Medical Press

The coronavirus was discovered after a lot of investment by China's People's Liberation Army (PLA) and was intentionally released from a lab as a bio-weapon, Chinese virologist Dr Le-Meng Yan said.

Yan, who was previously a post-doctoral fellow at Hong Kong University, fled to United States in April last year. In a paper published in September last year, she claimed that coronavirus was created by China in a research lab.

"...this document is one of the 'smoking gun' that can prove China has a long-term programme of non-traditional bio-weapons and (plans) to use it to conquer the whole world," Yan told India Today in an interview. She was referring to the leaked documents sourced by the US State Department which said Chinese military scientists investigated weaponising coronaviruses five years before the COVID-19 pandemic.

Yan said China's objective is to deny the allegations and use misinformation to mislead the world when people realise the virus has come from a lab.

Also read: Chinese military probed weaponising coronavirus in 2015, claim leaked documents

"I started telling people via YouTube anonymously from last January that this virus came from the PLA lab and that they discovered coronavirus after a lot of investment. They finally got a human-target virus and it was intentionally released. Also, the Chinese government knows it and that's why they immediately had a response after the waiting hours," she claimed.

Yan said the leaked documents show that genetic engineering of viruses in 2015 is not the beginning of China's study of contemporary bio-weapons, but is just one step in study of bio-weapons of which there is evidence now.

"But after that, they have also modified a lot and they have recruited a lot of labs under the cover of silo labs, under the cover of international labs, and working with the military to develop it. So they have better knowledge and experience after five-six years and that's what made COVID-19 happen," the virologist said.

US officials allegedly obtained papers, written in 2015 by military scientists and senior Chinese public health officials, as part of their own investigation into the origins of COVID-19.

In the papers, Chinese scientists described SARS coronaviruses - of which COVID is one example - as presenting a "new era of genetic weapons".

Also read: Another false hope in battle against Covid-19...What is Ivermectin?

Read more:
Coronavirus released intentionally, China using misinformation to mislead world: Chinese virologist - Business Today

This week, mosquito eggs placed in the Florida Keys are expected to hatch tens of thousands of genetically modified mosquitoes, a result of the first U.S. release of such insects in the wild. A biotechnology firm called Oxitec delivered the eggs in late April as part of a federally approved experiment to study the use of genetic engineeringrather than insecticidesto control disease-carrying mosquito populations. The move targets an invasive species, called Aedes aegypti, that carries Zika, dengue, chikungunya, yellow fever and other potentially deadly diseases, some of which are on the rise in Florida.

The experiment relies on a genetic alteration that will be lethal to a large number of future offspring. In this case, male mosquitoes have been modified to carry a gene that makes their female progeny dependent on the antibiotic tetracyclineand thus fated to die in the wild. As the mating cycle repeats over generations, female numbers are depleted, and the population is suppressed. The modified insects eventually die off, making this approach self-limiting.

Oxitec overcame significant regulatory hurdles before getting the go-ahead from the U.S. Food and Drug Administration in 2016 and then the Environmental Protection Agency in 2020. If the current pilot effort is successful, the firm is set to release as many as 20 million more males in the prime of Floridas mosquito season later this year. The results of the experiment could ultimately help address concerns about releasing genetically modified organisms into the wild.

To learn more about the risks and rewards of Floridas foray into bioengineered pest control, Scientific American spoke with Omar Akbari, a molecular biologist whose lab works on genetic control technologies at the University of California, San Diego. He is also a co-founder of Agragene, a biotech company that is using genetically engineered agricultural pests as a biological pest control.

[An edited transcript of the interview follows.]

Do you think the Aedes aegypti experiment in the Florida Keys will reduce the spread of mosquito-borne diseases?

The current method of controlling this species is to use insecticides, but they dont really work well. Weve noticed resistance in the field, so new technologies are definitely needed.

Oxitecs technology for releasing genetically modified insects has been tested in other places. [The company has] reported reaching A. aegypti population suppression of more than 90 percent in many of their releases, including effective control of the A. aegypti population in Brazil. Given its prior testing, the experiment in the Keys is likely to work and to suppress A. aegypti populations. And hopefully it will directly translate into an epidemiological impact, effectively reducing disease transmission.

How safe is this technology?

Its extremely safe. The EPA has done its due diligence and tested many of the potential side effects of this technology. The real question here is: What are the existing control mechanisms that are in place? This mosquito has been controlled using many different broad-spectrum insecticides in Florida, including pyrethroids that also kill honeybees, ladybugs, dragonflies and other insects. Pictures show aerial spraying of insecticides from airplanes over neighborhoods in Florida during the Zika virus outbreak in 2016. By comparison, Oxitecs technology is extremely safe. Its only going to target A. aegypti, and youre using the mosquito to control the mosquito.

Is there a risk to the ecosystem?

Its a misconception that this process could get rid of all mosquitoes. There are more than 3,500 different species of mosquitoes on earth. A handful of them transmit pathogens. Oxitec is not trying to eliminate all mosquitoes. [The company is] getting rid of one mosquito species from a localized population to stop it from transmitting pathogens to humans. And this mosquito speciesA. aegyptiis invasive and doesnt have a purpose in this environment. So I dont think there will be any negative environmental impact from removing the species from the environment.

Do you anticipate the future use of Oxitecs technology in other U.S. states?

Right now it is only approved to do mosquito egg releases in that one area of Florida. Its authorized here for experimental use. And the technology is localized. These mosquitoes cant travel very far.

The first requirement for use of the technology in other areas will be success with the current experiment in Florida. Once that is in hand, Oxitec can apply for more permits to do broader releases in other areas. If that were to happen, the process would resemble what took place in Florida. I think [Oxitec] would connect with the local mosquito-control districts in those locations and coordinate releases and monitoring the density of the A. aegypti female population over time. Getting approval in other locations might also require putting it on a ballot to get the public to weigh in on the decision, as was done in Florida.

What are the possible limitations of this approach to controlling mosquitoes that spread diseases?

One question is scalability. Can they scale this technology to eliminate this pest from, lets say, all the states in America that its present in, which is basically half of the U.S.? Or is it only useful in small communities? And if they scale if it, what is the cost associated with that?

Also, species-specific technology is a double-edged sword. On the one hand, youre only targeting one species. On the other hand, there are often multiple species transmitting a pathogen. For example, in Brazil, you have two different species that transmit dengue virusA. aegypti and Aedes albopictus. Thats also the case in Florida. So if you get rid of one of them, the other is still out there.

With global warming, how likely is it that other regions will take the same course that the Florida Keys mosquito district has?

Some already have. Oxitec has received approvals to do releases of its modified A. aegypti mosquitoes in the Cayman Islands and Panama. It is doing trials in Indiagenetically modified mosquitoes are released into cages with wild-type mosquitoes to mate and then compared with cages without the modified insect. [Others have] done releases in Malaysia and Australia. And as there are more examples of success stories, I think more countries will be willing to adopt this technology, assuming that the costs make sense.

With global warming, the habitable range of A. aegypti mosquitoes is expanding. The species now is present in many U.S. states, whereas 10 years ago it wasnt. This, too, is going to become more important as this mosquito species becomes more prevalent and the pathogens also become more prevalent.

What biological pest-control technologies are you currently working on?

Our lab has a [preprint] paper currently under review describing a new CRISPR-based technology that can be used to eliminate A. aegypti populations. Its also self-limiting. Were excited about this because we were able to eliminate the populations in experimental cages in the lab. And we think this technology might be a next-generation technology that can be used alongside the Oxitec technology. The outcome is very similar.

See the original post:
First Genetically Modified Mosquitoes Released in U.S. Are Hatching Now - Scientific American

SCOTLAND'S bloodthirsty midges may finally meet their match, as a result of revolutionary genetic manipulationtechniquesthat could stop the pesky insects biting chunks out of the tourist industry.

News that a British biotechnology company has used genetically engineeringto produce non-biting mosquitoes in Florida to help curb yellow and dengue fevers and ultimately malaria may spell long-term hope of beating the annual Scottish scourge.

4

Oxford-based Oxytec and American partners recently released genetically modified mosquito larvae in the Florida Keys after gene editing suppressed wild, disease-carrying populations without recourse to pesticides.

While midges share some characteristics with mosquitoes, Dr Simon Carpenter, a leading entomologist from the Pirbright Institute and part of the team that generated and built the first ever complete genome of a Culicoides biting midge, said using the same process in Scotland was more complex - and might take 20 years.

He toldThe Sunday Times: I would be amazed if no one took it on as a project. I would be surprised if there is not a genome (by then).

4

Professor Bruce Whitelaw, director of the Roslin Institute which gave the worldDolly the sheep, said it was conceptually possible to use the technology to reduce midge and even deer tick populations which can spread Lyme Disease.

But he added: [We] would need a lot of basic research to understand fertility in midges and establish the needed genetic engineering capability.

Carpenter said the sheer abundance of midges made it currently really difficult to knock out enough of the population to make a difference.

4

He added: Mosquitoes are pretty abundant in Florida but Culicoides Impunctatus - the species of midge native to Scotland - is insane.

He said that during research we were collecting 600 midges off an exposed arm.

During the controversial Florida project, 12,000 males were released but eventually hundreds of millions could be hatched.

4

The genetic material in the male mosquitoes was altered to pass on a lethal gene to female offspring, preventing them manufacturing an essential protein and reaching maturity. They then die off.

Whitelaw added: They try and mate but nothing will come of it."

"The insects only have to be released once and they drive through a population.

Oxytec claims the work is sustainable and eco-friendly and less damaging than toxic pesticides which insects can become resistant to.

Carpenter said researching midges was low down on the list of funding priorities because the end point of the bite is not fatal.

Deer tick were likely to be addressed ahead of midges, he added.

Swarms of midges descend across a quarter of the country every summer spreading viruses like blue-tongue disease in cattle and making life miserable for holidaymakers and locals.

It has been estimated to cost tourism 268million.

People rely on a variety of repellents to ward away midges.

Many walkers swear by Avons Skin So Soft and essential oils, while others resort to midge hoods and sophisticated machines that suck up huge quantities of the insects.

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Morag Phillips, who grew up on Skye and still visits the island, said: Certainly genetic modification would be good for tourism and make it easier for people to carry on working outside.

"The words genetic modification conjures up scary thoughts. But the idea of a midge that doesnt bite sounds like heaven to me.

But they are not universally detested. Adult midges are consumed by spiders and swallows and their larve are eaten by fish, dragonfly larvae, water beetles and other predators.

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We now live in a technologically advanced world that affects every aspect of our lives. Agriculture, for example, is an area where technology is fundamental. It has played a significant role in the growth of the agriculture industry, and the use of biotechnology has made it possible to grow crops in a desert. The plants have been designed to withstand extreme climatic conditions such as drought. As a result, technology is critical in agriculture, as it is in all other fields.

Furthermore, agriculture is the worlds most important field for producing food for humans. After the service sector, agriculture is the second-largest source of jobs, accounting for 28% of global employment. Agriculture employs about 1.3 billion people worldwide.

Keep in mind that farming is reliant on the natural world. Agriculture has been harmed in many areas of the world due to massive climate changes brought about by global warming. As a result, farmers today must rely on modern agriculture technology to make farming more efficient and affordable.

Every farmer should adopt modern farming technology to make agriculture more efficient while also lowering production and labor costs. Here are a few examples that might pique your interest.

Irrigation is arguably the most advanced in the agriculture sector when using technology to increase control and production. Cell phones and computers can now receive status reports on pivot results, soil moisture sensing, weather, and other field data, providing end-users with on-the-go tools to make and execute irrigation management decisions.

It refers to programming and coding that users can customize to meet their specific requirements. The most well-known open-source frameworks for creating an online presence are Drupal and WordPress, which create a blueprint that developers can then modify.

Small farms may use a variety of open-source services. Small farmers and homesteaders may use the farm management system to help record keeping, planning, and other management tasks. This form of technology aids small-scale farmers in maximizing their capacity to collect and distribute data in one location.

Genetic engineering and biotechnology are other terms for the process of enhancing the qualities of a seed. Reduced farm inputs imply that the farmer saves money on farm asset expenses as well. Among other things, modern farming technology aspires to achieve two primary goals. That is a more prosperous economy with a higher yield.

Some of the perspectives worth considering are how to apply and sort compost, water system, theatre, intensive culturing, monoculture, and the use of various assets. Regardless, farmers must grasp the concept of modern farming and the use of technology to achieve these goals.

Farm equipment has advanced significantly over time. The combine has supplanted the threshing machine, which is usually a self-propelled device that either picks up windrowed grain or cuts and threshes it in one move. A seed drill also simplifies the processing of sowing seeds and can effectively plant large areas of the crop. As a result, youll be able to get higher yields.

They are used for a variety of farming activities, including harvesting and weed control. Smart tractors and combines, for example, can operate autonomously and without the need for human intervention, allowing them to be used at any time of day. They may also include a variety of extra features, such as built-in pest control systems.

The sophistication of computerized weather modeling is growing. Farmers can use dedicated onboard and handheld farm technology and mobile applications that operate on almost any user device to access online weather services that are solely focused on agriculture. This technology will provide adequate warning of frost, hail, and other weather events so that farmers can take measures to protect their crops or, at the very least, reduce losses.

These devices can detect crop health as well as critical nitrogen levels in the water. The ground floor, soil characteristics such as ph level, the electrical conductivity of soil, and organic matter content are all measured using sensor technology. The sensors are both long-lasting and inexpensive. It protects the ecosystem on the farm by reducing fertilizer levels in lakes, minimizing erosion, and conserving water.

Net farm income has risen dramatically as a result of both of these developments. The key advantages of these systems are the efficient use of time, the reduction of resources, and real-time monitoring, both of which reduce farm management costs significantlyone of the methods that can assist you in reaping the benefits of farming.

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Recent technology has been developed to precisely engineer genes for traits of interest. This approach is known as CRISPR gene editing. Gene editing is distinct from other forms of genetic engineering, such as transgenic technology, often colloquially referred to as GMO. With gene editing, the final product can match that obtained by conventional plant breeding, but in a much shorter timeframe. CRISPR has been applied in many agronomic crops and is poised to make contributions in strawberry. We anticipate that, over the next decade, CRISPR and other gene editing techniques will be used to rapidly develop elite strawberry varieties with improved disease resistance, fruit quality, and other valuable attributes.What is CRISPR gene editing?

One of the great disappointments in the pursuit of improved varieties is the discovery of a new advanced selection that would be valued by the industry except for one critical flaw. Gene editing technology can be used to almost surgically adjust the gene or genes behind that deleterious trait. CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is one form of gene editing that can be used to precisely modify a gene of interest without otherwise compromising the favorable traits of an elite variety (Rani et al. 2016; Bortesi and Fischer 2015). For example, instead of breeding for many years to move a disease resistance gene from a wild strawberry into a modern strawberry, gene editing allows a direct introduction of the genetic information. Think of it as a cut and paste mechanism. This is particularly useful for cultivated strawberries because they are genetically complex, making conventional breeding difficult. The UF/IFAS strawberry breeding program has identified several important gene regions controlling disease resistance traits that are directly relevant to Florida growers. By using CRISPR technology, these genes or gene variants can be moved into desirable genetic backgrounds that can be further moved via conventional crossing in later generations. We can utilize established DNA marker-assisted breeding tools to track the edited genes in subsequent generations, adding to the speed of new variety development. Based on policy discussions, it is possible that the first-generation plants containing the edited genes will not require extensive regulation, and these tools will be extremely valuable in the long-term efforts of every strawberry breeding program.

Transgenic technology refers to the transfer of a genetic material from one species to another. CRISPR, on the other hand, can be used to precisely change DNA sequence, switching it from one naturally occurring variant to another naturally occurring variant. Using this new technique, we can cut a strawberrys genome at a desired location so that existing genes can be removed or added. Early indications suggest that gene editing should be regulated like conventionally bred crops, as the final product can simply match what may be done by conventional breeding. Recently, the USDA announced that it would not regulate a new mushroom developed using CRISPR (Waltz 2016). Countries like Sweden and Argentina have made similar proclamations, indicating that the finished CRISPR-edited varieties do not fall under certain regulations because they do not contain foreign DNA.

While the finished varieties do not contain DNA from other organisms, the process introduces genetic information that orchestrates the desired genetic change. The first step is to develop and optimize a tissue culture and transformation system (protocols to introduce foreign DNA to new plants) for UF strawberry lines, so that new plants can be regenerated from cells containing introduced DNA. However, just as each cultivar has different traits and qualities, they also behave differently with respect to introduction of new genes.

The recent cultivar Sweet Sensation Florida127 and advanced selection Florida Brilliance (FL 13.26-134) were used for tissue culture optimization. As shown in Figure 1, callus induction was tested with different strawberry tissues, and embryogenic callus growth was most vigorous on stolons (runners) and petioles. To identify the optimal conditions for shoot and root regeneration for Sweet Sensation Florida127 and Florida Brilliance (FL 13.26-134), explants were grown on a range of media with varying compositions of plant growth regulators.

About one inch of petiole or stolon from the leaf-end or shoot-end, respectively, were collected from greenhouse grown plants and used for the tissue culture process. Optimal conditions for tissue culture medium, nutrient, and hormone were tested for the UF accessions. The runner (stolon) produced calli more vigorously than petiole segments. It takes about 14 weeks for Sweet Sensation Florida127 and Florida Brilliance (FL 13.26-134) to develop from embryogenic callus to young plantlet in rooting media (Figure 2). Florida Brilliance (FL 13.26-134) produced more regenerated plants than Sweet Sensation Florida127.

Once the genetically engineered gene product is ready for CRISPR gene editing, transformation, where the new genetic material is delivered to a single strawberry cell, is the first step in the genetic engineering process. For DNA delivery, two major transformation methods, such as Agrobacterium tumefaciensmediated or biolistic (gene gun)mediated transformation, are widely used for CRISPR gene editing. Agrobacterium tumefaciens is a widespread, naturally occurring soil bacterium that causes crown gall in many plant species and has the ability to introduce new genetic material into plant cells (Gelvin 2003). This bacterium works as a natural genetic engineer and is used in labs for plant transformation. Gene-edited plants using Agrobacterium-mediated transformation will contain foreign bacterial DNA sequences. It is not an easy process to remove the bacterium-derived DNA sequences through breeding.

In contrast, we are currently using a biolistic particle bombardment method in which DNA-coated metal particles are delivered to the plant cells using a gene gun (Figure 3). This method can be applied to a wide range of cell and tissue types, and there is no need for bacterial DNA insertion. Later, the gene edited tissues can be regenerated to mature plants using the tissue culture protocol outlined above.

Recently, the development of gene editing using protoplasts and regeneration of plants from protoplasts has been demonstrated in other plant species (Woo et al. 2015; Kanchiswamy 2016; Dutt et al. 2015). This method is known as a completely DNA-free gene editing system. Briefly, the protein/genetic material complex for gene editing will be assembled in vitro and the complex mixed with strawberry protoplast isolated from embryogenic calli and polyethylene glycol, which allows direct transfer by endocytosis into protoplasts. The gene edited with protoplasts is cultured (cell suspension culture) into calli, and mature plants can be regenerated using the tissue culture protocol outlined above. We are currently developing this cell suspension culture protocol at UF.

The UF strawberry breeding program provides a direct commercialization path for disease-resistant varieties to reach Florida growers. New varieties with better resistance will bring increased profitability to the Florida strawberry industry. The Florida Strawberry Growers Association estimates that diseases cost the Florida industry at least $15 million each year. In the last two years, the UF strawberry breeding program has identified regions of strawberry chromosomes that control resistance to bacterial angular leaf spot (Roach et al. 2016) and Phytophthora crown rot (Mangandi et al. 2017), with more to come for other diseases such as Colletotrichum crown rot, charcoal rot, and anthracnose fruit rot. Efforts are ongoing to narrow these chromosomal regions down and identify the exact gene sequences that provide these disease resistances, with Phytophthora resistance as the first priority. Our goal is to add Phytophthora resistance to Florida Brilliance (FL 13.26-134) and Sweet Sensation Florida127, which are currently highly susceptible to this disease. Evaluations of the gene-edited lines will be performed in concert with crosses to integrate the changes into other major varieties and advanced selections with conventional hybridization.

Bortesi, L. and R. Fischer. 2015. The CRISPR/Cas9 system for plant genome editing and beyond. Biotechnol Adv. 33(1): 4152.

Dutt, M. et al. 2015. Transgenic Citrus Expressing an Arabidopsis NPR1 Gene Exhibit Enhanced Resistance against Huanglongbing (HLB; Citrus Greening). PLoS One 10(9):e0137134.

Gelvin, S.B. 2003. Agrobacterium-mediated plant trans-formation: the biology behind the gene-jockeying tool. Microbiol Mol Biol Rev. 67(1):1637.

Kanchiswamy, C.N. 2016. DNA-free genome editing meth-ods for targeted crop improvement. Plant Cell Rep. 35(7): 14691474.

Mangandi, J. et al. 2017. Pedigree-Based Analysis in a Mul-tiparental Population of Octoploid Strawberry Reveals QTL Alleles Conferring Resistance to Phytophthora cactorum. G3 7(6): 17071719.

Rani, R., et al. 2016. CRISPR/Cas9: a promising way to exploit genetic variation in plants. Biotechnol Lett.

Roach, J.A., et al. 2016. FaRXf1: a locus conferring resistance to angular leaf spot caused by Xanthomonas fragariae in octoploid strawberry. Theor Appl Genet.

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Waltz, E. 2016. Gene-edited CRISPR mushroom escapes US regulation. Nature 532(7599): 293.

Woo, J.W., et al. 2015. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat Biotechnol. 33(11): 11621164.

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Tired of eating strawberries that taste like cardboard? CRISPR gene editing poised to improve 'fruit quality' and disease resistance of...