Daily Archives: June 26, 2022

by Dr. Surendra K Chikara

Everyones DNA is as unique as their fingerprint. Information about a persons genes, environment and lifestyle factors can be used to prevent or manage disease and this is usually referred to as precision medicine. This form of personalized healthcare has been around for a few years and has been increasingly gaining popularity. Thanks to the Human Genome Project (1990-2003) that sequenced around 20,000 genes and created a breakthrough in healthcare. We are now able to use genomics to predict, prevent and manage disease better than ever.

As of today, we are experiencing a revolutionary shift towards precision medicine. We can quickly sequence DNA at a large scale and help thousands of people manage and prevent disease. Genomics has been especially valuable for identifying rare genetic diseases that had previously taken years to diagnose, ending uncertainty and suffering for many people.

In addition to reducing the risk of disease, genomic testing can serve as a source of data. Many organizations are now investing time and money into building databases of genetic biomarkers for various chronic diseases, so they can be identified early. Such databases could deliver a definitive diagnosis in seconds which could significantly bring down treatment costs at a global level.

The use of genomics in the healthcare industry has given a new perspective on managing disease; people are now focused on preventing disease rather than curing it. There are numerous start-ups that are working on utilizing genomic testing to prevent disease by helping people find the root cause of their health issues.

The need for preventive healthcareA World Economic Forum study estimates that the global economic impact of cancer, diabetes, mental illness, heart disease, and respiratory disease could reach USD 47 trillion over the next 20 years. The increased demand on healthcare systems could be lessened through breakthroughs like genomic testing because they make it possible to prevent disease.The growing prevalence of chronic diseases and demand for personalized medicine have contributed to the precision medicine market size to grow from USD 8.2 billion to 16.4 billion by 2025.

The Pandemic further proved to be an eye-opener for many people, especially those with chronic diseases, since they were prone to a higher risk for catching the virus and developing complications. This highlighted the immense need for preventing such diseases in the first place and further emphasized the importance of the need for preventive healthcare in the country.

Preventive healthcare is the future Genomics has created a shift in peoples mindset towards disease prevention. It has become increasingly clear in recent years that genomic testing and precision medicine is the wave of the future. Just like health insurance has now become commonplace in India, genomic testing would soon become the first line of defense against chronic disease. It is only a matter of time before genomic testing and thus precision medicine enters the mainstream of healthcare in India as well.

Dr. Surendra K Chikara - Founder & CEO - Bione

(DISCLAIMER: The views expressed are solely of the author and ETHealthworld does not necessarily subscribe to it. ETHealthworld.com shall not be responsible for any damage caused to any person / organisation directly or indirectly.)

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Genomics: A Revolution in Health Care? - ETHealthWorld

An appealing alternative to the Streptococcus pyogenes CRISPR nuclease SpyCas9, are Type V CRISPR Cas12a nucleases, commonly isolated from Acidaminococcus (Asp) and Lachnospiraceae (Lba). These Cas12a nucleases embody several desirable attributes that SpyCas9 lacks: they exhibit greater editing precision, recognize a thymine-rich PAM (protospacer adjacent motifa two-to-six base sequence following the nuclease target), use a single CRISPR-RNA to detect its target, cut DNA in a staggered fashion generating overhangs, process CRISPR arrays, and have been shown to function in diverse organisms ranging from plants to mammals. However, Cas12a nucleases exhibit lower editing rates than SpyCas9 in primary cells.

In a study published in GENmagazines sister journal,GEN Biotechnology (Optimization of Nuclear Localization Signal Composition Improves CRISPR-Cas12a Editing Rates in Human Primary Cells), Scot Wolfe, PhD, professor of molecular, cell and cancer biology at the University of Massachusetts Chan Medical School and his team, increased Cas12as on-target gene editing rate to nearly 100% by engineering the configuration of the enzymes nuclear localization signal (NLS). These advancements to the Cas12a editing framework could improve the use of this nuclease to uncover functions of new genes and develop new CRISPR-based treatments.

Previous work by our laboratories and others indicated that the efficiency of Cas12a editing in CD34+ hematopoietic stem and progenitor cells could potentially be improved by increasing the efficiency of its nuclear import, said Wolfe.

In earlier studies, Wolfes team had enhanced SpyCas9 gene editing in primary cells by optimizing the NLS sequence composition and number. They had found adding one NLS at the amino-terminus and two at the carboxy-terminus of the nuclease markedly improved SpyCas9s (3xNLS-SpyCas9) editing efficiency in hematopoietic stem and progenitor cells (HSPCs). They had then added two NLSs to the carboxy-terminus of Cas12a but did not achieve the same efficiency of targeted mutagenesis as the engineered SpyCas9 with three NLSs.

Ben Kleinstiver, PhD, assistant professor of pathology at Massachusetts General Hospital and Harvard Medical School, said, Genome editing efficiency is impacted by many different variables, including the concentration of a CRISPR-Cas enzyme in the nucleus where it performs its function. Researchers have previously dedicated substantial effort to improve CRISPR nuclease expression and nuclear localization for SpyCas9, but comparatively fewer optimizations have been performed for Cas12a. (Kleinstiver was not involved in the current study).

In the current study, Wolfes team developed three NLS C-terminus variants of Cas12a where they substituted the previously used simian virus NLS (SV40) with a more efficient NLS of a proto-oncogene (c-Myc). In addition, they added a third NLS to the carboxy end to achieve an editing platform at par with 3xNLS-SpyCas9 in editing efficiency. The researchers observed increased knockout efficiency in all three Cas12a orthologs (Asp, Lba, and engineered-Asp) they tested, which suggests this triple NLS strategy could be effective in improving the activity of other members of the Cas12a family, without decreasing the enzymes inherent specificity.

The study used standard electroporation to deliver the engineered Cas12a ribonucleoproteins (RNPs) into transformed human cells lines (HEK293T, Jurkat, and K562 cells) and into primary cells (natural killer cells and CD34+ HSPCs) to improve indel frequencies.

We believe that the improved NLS sequence architecture described in this paper will increase the efficiency of genome editing by Cas12a in primary cells, thus leading to increased levels of therapeutic genome editing in a variety of applications, said Wolfe. The researchers claim this strategy of enhancing the NLS sequence can be widely applied to other Cas12a orthologs and variants with similar outcomes.

The Wolfe lab and collaborators had previously demonstrated increased activity with a new NLS framework for SpyCas, so it is exciting that they demonstrated success with a new NLS for Cas12a in this publication. It is important to have additional NLSs to test in the growing list of nucleases and cell types, said Thomas Cradick, PhD, CSO at Excision BioTherapeutics. (Cradick was not involved in the current study.)

Kleinstiver said, Luk et al., demonstrated that the efficiency of editing with various Cas12a enzymes can be improved by using a more optimal configuration of NLSs. The effect of this optimization was most striking in lipid-based transfections (nucleofections) in transformed cell lines, with a more modest improvement in primary cells, the latter of which due to already high levels of editing in primary cells.

This study resurfaces a really important consideration, that you can only edit cells as efficiently as your enzyme is designed to. There are lots of knobs to turn to optimize and improve editing efficiency, and the NLS architecture clearly plays a key role in regulating the nuclear concentration, and thus the potency, of the editor, added Kleinstiver.

Nicole Gaudelli, PhD, director and head of gene editing platform technologies at Beam Therapeutics, who was not involved in the current study, said, In addition to advancing Cas12a gene editing applications, these learnings may potentially be evaluated for other gene editing tools to further increase editing efficiencies and provide greater therapeutic benefit if higher levels of gene correction or modification can be achieved.

This study was rigorously done in multiple cell types that show the robustness of the data. I liked how they delivered Cas12 as an RNP, as this is therapeutically relevant and greatly reduces off-target editing, said Alexis Komor, PhD, assistant professor of chemistry and biochemistry at the University of California, San Diego, who was not involved in the study.

I also liked this work because it uses a very universal approach to improve editing (the modifications they made to the system can be applied to any genome editing agent), and they demonstrated its utility with multiple Cas12 enzymes (which have slightly different PAMs, which is nice). Overall, its a useful and practical study, Komor continued.

As we continue the deployment of diverse CRISPR-Cas effectors in the clinic, it is important to individually engineer each molecular machine for optimal efficiency and specificity. Here, the authors show how NLS can be optimized for enhanced activity in medically relevant human primary cells, said Rodolphe Barrangou, PhD, professor of food, bioprocessing, and nutrition at North Carolina State University (NCSU), editor-in-chief of The CRISPR Journal, and CEO of TreeCo, a company that uses CRISPR to produce genetically enhanced trees. Barrangou was not part of the current study.

Optimizing on-target mutagenesis rates whilst maintaining specificity is key for successful translation to the clinic, reaffirmed Jennifer Harbottle, PhD, a senior scientist at Horizon Discovery, who was not part of this study. The Cas12a NLS variant developed by Scot Wolfes lab holds the potential to lower dosage whilst exerting therapeutic effect.

It will be of interest to see this strategy expanded to other Type V systems, and track efficiency of delivery in a wider range of cell types and tissues, added Harbottle. Comprehensively evaluating the genomic integrity of edited cells, particularly the occurrence of structural variants and chromosomal rearrangements compared to editing by canonical Cas9 systems, will be critical to push the optimized Type V variants towards in vivo use in humans.

In future studies, Wolfe intends to continue refining Cas12a nucleases to edit specific therapeutic targets. He said, We are particularly interested in applications for certain hematopoietic disorders and muscular dystrophies.

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CRISPR-Cas12a Editing Rates Improve with Better Directions to the Nucleus - Genetic Engineering & Biotechnology News

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Applications for the CAD software extend far beyond medicine and throughout the burgeoning field of synthetic biology, which involves redesigning organisms to give them new abilities. For example, we envision users designing solutions for biomanufacturing; it's possible that society could reduce its reliance on petroleum thanks to microorganisms that produce valuable chemicals and materials. And to aid the fight against climate change, users could design microorganisms that ingest and lock up carbon, thus reducing atmospheric carbon dioxide (the main driver of global warming).

Our consortium, GP-write, can be understood as a sequel to the Human Genome Project, in which scientists first learned how to "read" the entire genetic sequence of human beings. GP-write aims to take the next step in genetic literacy by enabling the routine "writing" of entire genomes, each with tens of thousands of different variations. As genome writing and editing becomes more accessible, biosafety is a top priority. We're building safeguards into our system from the start to ensure that the platform isn't used to craft dangerous or pathogenic sequences.

Need a quick refresher on genetic engineering? It starts with DNA, the double-stranded molecule that encodes the instructions for all life on our planet. DNA is composed of four types of nitrogen basesadenine (A), thymine (T), guanine (G), and cytosine (C)and the sequence of those bases determines the biological instructions in the DNA. Those bases pair up to create what look like the rungs of a long and twisted ladder. The human genome (meaning the entire DNA sequence in each human cell) is composed of approximately 3 billion base-pairs. Within the genome are sections of DNA called genes, many of which code for the production of proteins; there are more than 20,000 genes in the human genome.

The Human Genome Project, which produced the first draft of a human genome in 2000, took more than a decade and cost about $2.7 billion in total. Today, an individual's genome can be sequenced in a day for $600, with some predicting that the $100 genome is not far behind. The ease of genome sequencing has transformed both basic biological research and nearly all areas of medicine. For example, doctors have been able to precisely identify genomic variants that are correlated with certain types of cancer, helping them to establish screening regimens for early detection. However, the process of identifying and understanding variants that cause disease and developing targeted therapeutics is still in its infancy and remains a defining challenge.

Until now, genetic editing has been a matter of changing one or two genes within a massive genome; sophisticated techniques like CRISPR can create targeted edits, but at a small scale. And although many software packages exist to help with gene editing and synthesis, the scope of those software algorithms is limited to single or few gene edits. Our CAD program will be the first to enable editing and design at genome-scale, allowing users to change thousands of genes, and it will operate with a degree of abstraction and automation that allows designers to think about the big picture. As users create new genome variants and study the results in cells, each variant's traits and characteristics (called its phenotype) can be noted and added to the platform's libraries. Such a shared database could vastly speed up research on complex diseases.

What's more, current genomic design software requires human experts to predict the effect of edits. In a future version, GP-write's software will include predictions of phenotype to help scientists understand if their edits will have the desired effect. All the experimental data generated by users can feed into a machine-learning program, improving its predictions in a virtuous cycle. As more researchers leverage the CAD platform and share data (the open-source platform will be freely available to academia), its predictive power will be enhanced and refined.

Our first version of the CAD software will feature a user-friendly graphical interface enabling researchers to upload a species' genome, make thousands of edits throughout the genome, and output a file that can go directly to a DNA synthesis company for manufacture. The platform will also enable design sharing, an important feature in the collaborative efforts required for large-scale genome-writing initiatives.

There are clear parallels between CAD programs for electronic and genome design. To make a gadget with four transistors, you wouldn't need the help of a computer. But today's systems may have billions of transistors and other components, and designing them would be impossible without design-automation software. Likewise, designing just a snippet of DNA can be a manual process. But sophisticated genomic designwith thousands to tens of thousands of edits across a genomeis simply not feasible without something like the CAD program we're developing. Users must be able to input high-level directives that are executed across the genome in a matter of seconds.

Our CAD program will be the first to enable editing at genome-scale, with a degree of abstraction and automation that allows designers to think about the big picture.

A good CAD program for electronics includes certain design rules to prevent a user from spending a lot of time on a design, only to discover that it can't be built. For example, a good program won't let the user put down transistors in patterns that can't be manufactured or put in a logic that doesn't make sense. We want the same sort of design-for-manufacture rules for our genomic CAD program. Ultimately, our system will alert users if they're creating sequences that can't be manufactured by synthesis companies, which currently have limitations such as trouble with certain repetitive DNA sequences. It will also inform users if their biological logic is faulty; for example, if the gene sequence they added to code for the production of a protein won't work, because they've mistakenly included a "stop production" signal halfway through.

But other aspects of our enterprise seem unique. For one thing, our users may import huge files containing billions of base-pairs. The genome of the Polychaos dubium, a freshwater amoeboid, clocks in at 670 billion base-pairsthat's over 200 times larger than the human genome! As our CAD program will be hosted on the cloud and run on any Internet browser, we need to think about efficiency in the user experience. We don't want a user to click the "save" button and then wait ten minutes for results. We may employ the technique of lazy loading, in which the program only uploads the portion of the genome that the user is working on, or implement other tricks with caching.

Getting a DNA sequence into the CAD program is just the first step, because the sequence, on its own, doesn't tell you much. What's needed is another layer of annotation to indicate the structure and function of that sequence. For example, a gene that codes for the production of a protein is composed of three regions: the promoter that turns the gene on, the coding region that contains instructions for synthesizing RNA (the next step in protein production), and the termination sequence that indicates the end of the gene. Within the coding region, there are "exons," which are directly translated into the amino acids that make up proteins and "introns," intervening sequences of nucleotides that are removed during the process of gene expression. There are existing standards for this annotation that we want to improve on, so our standardized interface language will be readily interpretable by people all over the world.

The CAD program from GP-write will enable users to apply high-level directives to edit a genome, including inserting, deleting, modifying, and replacing certain parts of the sequence. GP-write

Once a user imports the genome, the editing engine will enable the user to make changes throughout the genome. Right now, we're exploring different ways to efficiently make these changes and keep track of them. One idea is an approach we call genome algebra, which is analogous to the algebra we all learned in school. In mathematics, if you want to get from the number 1 to the number 10, there are infinite ways to do it. You could add 1 million and then subtract almost all of it, or you could get there by repeatedly adding tiny amounts. In algebra, you have a set of operations, costs for each of those operations, and tools that help organize everything.

In genome algebra, we have four operations: we can insert, delete, invert, or edit sequences of nucleotides. The CAD program can execute these operations based on certain rules of genomics, without the user having to get into the details. Similar to the "PEMDAS rule" that defines the order of operations in arithmetic, the genome editing engine must order the user's operations correctly to get the desired outcome. The software could also compare sequences against each other, essentially checking their math to determine similarities and differences in the resulting genomes.

In a later version of the software, we'll also have algorithms that advise users on how best to create the genomes they have in mind. Some altered genomes can most efficiently be produced by creating the DNA sequence from scratch, while others are more suited to large-scale edits of an existing genome. Users will be able to input their design objectives and get recommendations on whether to use a synthesis or editing strategyor a combination of the two.

Users can import any genome (here, the E. coli bacteria genome), and create many edited versions; the CAD program will automatically annotate each version to show the changes made. GP-write

Our goal is to make the CAD program a "one-stop shop" for users, with the help of the members of our Industry Advisory Board: Agilent Technologies, a global leader in life sciences, diagnostics and applied chemical markets; the DNA synthesis companies Ansa Biotechnologies, DNA Script, and Twist Bioscience; and the gene editing automation companies Inscripta and Lattice Automation. (Lattice was founded by coauthor Douglas Densmore). We are also partnering with biofoudries such as the Edinburgh Genome Foundry that can take synthetic DNA fragments, assemble them, and validate them before the genome is sent to a lab for testing in cells.

Users can most readily benefit from our connections to DNA synthesis companies; when possible, we'll use these companies' APIs to allow CAD users to place orders and send their sequences off to be synthesized. (In the case of DNA Script, when a user places an order it would be quickly printed on the company's DNA printers; some dedicated users might even buy their own printers for more rapid turnaround.) In the future, we'd like to make the ordering step even more user-friendly by suggesting the company best suited to the manufacture of a particular sequence, or perhaps by creating a marketplace where the user can see prices from multiple manufacturers, the way people do on airfare sites.

We've recently added two new members to our Industrial Advisory Board, each of which brings interesting new capabilities to our users. Catalog Technologies is the first commercially viable platform to use synthetic DNA for massive digital storage and computation, and could eventually help users store vast amounts of genomic data generated on GP-write software. The other new board member is SOSV's IndieBio, the leader in biotech startup development. It will work with GP-write to select, fund, and launch companies advancing genome-writing science from IndieBio's New York office. Naturally, all those startups will have access to our CAD software.

We're motivated by a desire to make genome editing and synthesis more accessible than ever before. Imagine if high-school kids who don't have access to a wet lab could find their way to genetic research via a computer in their school library; this scenario could enable outreach to future genome design engineers and could lead to a more diverse workforce. Our CAD program could also entice people with engineering or computational backgroundsbut with no knowledge of biologyto contribute their skills to genetic research.

Because of this new level of accessibility, biosafety is a top priority. We're planning to build several different levels of safety checks into our system. There will be user authentication, so we'll know who's using our technology. We'll have biosecurity checks upon the import and export of any sequence, basing our "prohibited" list on the standards devised by the International Gene Synthesis Consortium (IGSC), and updated in accordance with their evolving database of pathogens and potentially dangerous sequences. In addition to hard checkpoints that prevent a user from moving forward with something dangerous, we may also develop a softer system of warnings.

Imagine if high-school kids who don't have access to a lab could find their way to genetic research via a computer in their school library.

We'll also keep a permanent record of redesigned genomes for tracing and tracking purposes. This record will serve as a unique identifier for each new genome and will enable proper attribution to further encourage sharing and collaboration. The goal is to create a broadly accessible resource for researchers, philanthropies, pharmaceutical companies, and funders to share their designs and lessons learned, helping all of them identify fruitful pathways for advancing R&D on genetic diseases and environmental health. We believe that the authentication of users and annotated tracking of their designs will serve two complementary goals: It will enhance biosecurity while also engendering a safer environment for collaborative exchange by creating a record for attribution.

One project that will put the CAD program to the test is a grand challenge adopted by GP-write, the Ultra-Safe Cell Project. This effort, led by coauthor Farren Isaacs and Harvard professor George Church, aims to create a human cell line that is resistant to viral infection. Such virus-resistant cells could be a huge boon to the biomanufacturing and pharmaceutical industry by enabling the production of more robust and stable products, potentially driving down the cost of biomanufacturing and passing along the savings to patients.

The Ultra-Safe Cell Project relies on a technique called recoding. To build proteins, cells use combinations of three DNA bases, called codons, to code for each amino acid building block. For example, the triplet 'GGC' represents the amino acid glycine, TTA represents leucine, GTC represents valine, and so on. Because there are 64 possible codons but only 20 amino acids, many of the codons are redundant. For example, four different codons can code for glycine: GGT, GGC, GGA, and GGG. If you replaced a redundant codon in all genes (or 'recode' the genes), the human cell could still make all of its proteins. But viruseswhose genes would still include the redundant codons and which rely on the host cell to replicatewould not be able to translate their genes into proteins. Think of a key that no longer fits into the lock; viruses trying to replicate would be unable to do so in the cells' machinery, rendering the recoded cells virus-resistant.

This concept of recoding for viral resistance has already been demonstrated. Isaacs, Church, and their colleagues reported in a 2013 paper in Science that, by removing all 321 instances of a single codon from the genome of the E. coli bacterium, they could impart resistance to viruses which use that codon. But the ultra-safe cell line requires edits on a much grander scale. We estimate that it would entail thousands to tens of thousands of edits across the human genome (for example, removing specific redundant codons from all 20,000 human genes). Such an ambitious undertaking can only be achieved with the help of the CAD program, which can automate much of the drudge work and let researchers focus on high-level design.

The famed physicist Richard Feynman once said, "What I cannot create, I do not understand." With our CAD program, we hope geneticists become creators who understand life on an entirely new level.

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DURHAM, N.C.--(BUSINESS WIRE)--Precision BioSciences, Inc. (Nasdaq: DTIL), a clinical stage gene editing company developing ARCUS-based ex vivo allogeneic CAR T and in vivo gene editing therapies, today announced it has entered into an exclusive worldwide in vivo gene editing research and development collaboration and license agreement with Novartis Pharma AG (the Agreement). As part of the Agreement, Precision will develop a custom ARCUS nuclease that will be designed to insert, in vivo, a therapeutic transgene at a safe harbor location in the genome as a potential one-time transformative treatment option for diseases including certain hemoglobinopathies such as sickle cell disease and beta thalassemia.

Under the terms of the Agreement, Precision will develop an ARCUS nuclease and conduct in vitro characterization, with Novartis then assuming responsibility for all subsequent research, development, manufacturing and commercialization activities. Novartis will receive an exclusive license to the custom ARCUS nuclease developed by Precision for Novartis to further develop as a potential in vivo treatment option for sickle cell disease and beta thalassemia. Precision will receive an upfront payment of $75 million and is eligible to receive up to an aggregate amount of approximately $1.4 billion in additional payments for future milestones. Precision is also eligible to receive certain research funding and, should Novartis successfully commercialize a therapy from the collaboration, tiered royalties ranging from the mid-single digits to low-double digits on product sales.

We are excited to collaborate with Novartis to bring together the precision and versatility of ARCUS genome editing with Novartis gene therapy expertise and commitment to developing one-time, potentially transformative treatment for hard-to-treat inherited blood disorders, said Michael Amoroso, Chief Executive Officer at Precision BioSciences. This collaboration will build on the unique gene insertion capabilities of ARCUS and illustrates its utility as a premium genome editing platform for potential in vivo drug development. With this Agreement, Precision, either alone or with world-class partners, will have active in vivo gene editing programs for targeted gene insertion and gene deletions in hematopoietic stem cells, liver, muscle and the central nervous system showcasing the distinctive versatility of ARCUS.

We identify here a collaborative opportunity to imagine a unique therapeutic option for patients with hemoglobinopathies, such as sickle cell disease and beta thalassemia a potential one-time treatment administered directly to the patient that would overcome many of the hurdles present today with other therapeutic technologies, said Jay Bradner, President of the Novartis Institutes for Biomedical Research (NIBR), the Novartis innovation engine. We look forward to working with Precision and leveraging the ARCUS technology platform, which could bring a differentiated approach to the treatment of patients with hemoglobinopathies."

The in vivo gene editing approach that we are pursuing for sickle cell disease could have a number of significant advantages over other ex vivo gene therapies currently in development, said Derek Jantz, Ph.D., Chief Scientific Officer and Co-Founder of Precision BioSciences. Perhaps most importantly, it could open the door to treating patients in geographies where stem cell transplant is not a realistic option. We believe that the unique characteristics of the ARCUS platform, particularly its ability to target gene insertion with high efficiency, make it the ideal choice for this project, and we look forward to working with our partners at Novartis to bring this novel therapy to patients.

Upon completion of the transaction, Precision expects that existing cash and cash equivalents, expected operational receipts, and available credit will be sufficient to fund its operating expenses and capital expenditure requirements into Q2 2024.

Precision BioSciences Conference Call and Webcast Information

Precision's management team will host a conference call and webcast tomorrow, June 22, 2022, at 8:00 AM ET to discuss the collaboration. The dial-in conference call numbers for domestic and international callers are (866)-996-7202 and (270)-215-9609, respectively. The conference ID number for the call is 6252688. Participants may access the live webcast on Precision's website https://investor.precisionbiosciences.com/events-and-presentations in the Investors page under Events and Presentations. An archived replay of the webcast will be available on Precision's website.

About ARCUS and Safe harbor ARCUS Nucleases

ARCUS is a proprietary genome editing technology discovered and developed by scientists at Precision BioSciences. It uses sequence-specific DNA-cutting enzymes, or nucleases, that are designed to either insert (knock-in), remove (knock-out), or repair DNA of living cells and organisms. ARCUS is based on a naturally occurring genome editing enzyme, I-CreI, that evolved in the algae Chlamydomonas reinhardtii to make highly specific cuts in cellular DNA. Precision's platform and products are protected by a comprehensive portfolio including nearly 100 patents to date.

Precision can use an ARCUS nuclease to add a healthy copy of a gene (or payload) to a persons genome. The healthy copy of the gene can be inserted at its usual site within the genome, replacing the mutated, disease-causing copy. Alternatively, an ARCUS nuclease can be used to insert a healthy copy of the gene at another site within the genome called a safe harbor that enables production of the healthy gene product without otherwise affecting the patients DNA of gene expression patterns.

About Sickle Cell Disease and Beta Thalassemia

Sickle cell disease (SCD) is a complex genetic disorder that affects the structure and function of hemoglobin, reduces the ability of red blood cells to transport oxygen efficiently and, early on, progresses to a chronic vascular disease.1-4 The disease can lead to acute episodes of pain known as sickle cell pain crises, or vaso-occlusive crises, as well as life-threatening complications.5-7 The condition affects 20 million people worldwide.8 Approximately 80% of individuals with SCD globally live in sub-Saharan Africa and it is estimated that approximately 1,000 children in Africa are born with SCD every day and more than half will die before they reach five.9,10 SCD is also a multisystem disorder and the most common genetic disease in the United States, affecting 1 in 500 African Americans. About 1 in 12 African Americans carry the autosomal recessive mutation, and approximately 300,000 infants are born with sickle cell anemia annually.11 Even with todays best available care, SCD continues to drive premature deaths and disability as this lifelong illness often takes an extreme emotional, physical, and financial toll on patients and their families.12,13

Beta thalassemia is also an inherited blood disorder characterized by reduced levels of functional hemoglobin.14 The condition has three main forms minor, intermedia and major, which indicate the severity of the disease.14 While the symptoms and severity of beta thalassemia varies greatly from one person to another, a beta thalassemia major diagnosis is usually made during the first two years of life and individuals require regular blood transfusions and lifelong medical care to survive.14 Though the disorder is relatively rare in the United States, it is one of the most common autosomal recessive disorders in the world.14 The incidence of symptomatic cases is estimated to be approximately 1 in 100,000 individuals in the general population.14, 15 The frequency of beta-thalassemia mutations varies by regions of the world with the highest prevalence in the Mediterranean, the Middle-East, and Southeast and Central Asia. Approximately 68,000 children are born with beta-thalassemia.16

About Precision BioSciences, Inc.

Precision BioSciences, Inc. is a clinical stage biotechnology company dedicated to improving life (DTIL) with its novel and proprietary ARCUS genome editing platform. ARCUS is a highly precise and versatile genome editing platform that was designed with therapeutic safety, delivery, and control in mind. Using ARCUS, the Companys pipeline consists of multiple ex vivo off-the-shelf CAR T immunotherapy clinical candidates and several in vivo gene editing candidates designed to cure genetic and infectious diseases where no adequate treatments exist. For more information about Precision BioSciences, please visit http://www.precisionbiosciences.com.

Forward-Looking Statements

This press release contains forward-looking statements, as may any related presentations, within the meaning of the Private Securities Litigation Reform Act of 1995. All statements contained in this herein and in any related presentation that do not relate to matters of historical fact should be considered forward-looking statements, including, without limitation, statements regarding the goal of providing a one time, potentially curative treatment for certain hemoglobinopathies, the success of the collaboration with Novartis, including the receipt of any milestone, royalty, or other payments pursuant to and the satisfaction of obligations under the Agreement, clinical and regulatory development and expected efficacy and benefit of our platform and product candidates, expectations about our operational initiatives and business strategy, expectations about achievement of key milestones, and expected cash runway. In some cases, you can identify forward-looking statements by terms such as aim, anticipate, approach, believe, contemplate, could, estimate, expect, goal, intend, look, may, mission, plan, potential, predict, project, should, target, will, would, or the negative thereof and similar words and expressions. Forward-looking statements are based on managements current expectations, beliefs and assumptions and on information currently available to us. Such statements are subject to a number of known and unknown risks, uncertainties and assumptions, and actual results may differ materially from those expressed or implied in the forward-looking statements due to various important factors, including, but not limited to: our ability to become profitable; our ability to procure sufficient funding and requirements under our current debt instruments and effects of restrictions thereunder; risks associated with raising additional capital; our operating expenses and our ability to predict what those expenses will be; our limited operating history; the success of our programs and product candidates in which we expend our resources; our limited ability or inability to assess the safety and efficacy of our product candidates; our dependence on our ARCUS technology; the initiation, cost, timing, progress, achievement of milestones and results of research and development activities, preclinical studies and clinical trials; public perception about genome editing technology and its applications; competition in the genome editing, biopharmaceutical, and biotechnology fields; our or our collaborators ability to identify, develop and commercialize product candidates; pending and potential liability lawsuits and penalties against us or our collaborators related to our technology and our product candidates; the U.S. and foreign regulatory landscape applicable to our and our collaborators development of product candidates; our or our collaborators ability to obtain and maintain regulatory approval of our product candidates, and any related restrictions, limitations and/or warnings in the label of an approved product candidate; our or our collaborators ability to advance product candidates into, and successfully design, implement and complete, clinical or field trials; potential manufacturing problems associated with the development or commercialization of any of our product candidates; our ability to obtain an adequate supply of T cells from qualified donors; our ability to achieve our anticipated operating efficiencies at our manufacturing facility; delays or difficulties in our and our collaborators ability to enroll patients; changes in interim top-line and initial data that we announce or publish; if our product candidates do not work as intended or cause undesirable side effects; risks associated with applicable healthcare, data protection, privacy and security regulations and our compliance therewith; the rate and degree of market acceptance of any of our product candidates; the success of our existing collaboration agreements, and our ability to enter into new collaboration arrangements; our current and future relationships with and reliance on third parties including suppliers and manufacturers; our ability to obtain and maintain intellectual property protection for our technology and any of our product candidates; potential litigation relating to infringement or misappropriation of intellectual property rights; our ability to effectively manage the growth of our operations; our ability to attract, retain, and motivate key executives and personnel; market and economic conditions; effects of system failures and security breaches; effects of natural and manmade disasters, public health emergencies and other natural catastrophic events; effects of COVID-19 pandemic and variants thereof, or any pandemic, epidemic or outbreak of an infectious disease; insurance expenses and exposure to uninsured liabilities; effects of tax rules; risks related to ownership of our common stock and other important factors discussed under the caption Risk Factors in our Quarterly Report on Form 10-Q for the quarterly period ended March 31, 2022, as any such factors may be updated from time to time in our other filings with the SEC, which are accessible on the SECs website at http://www.sec.gov and the Investors page of our website under SEC Filings at investor.precisionbiosciences.com.

References

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Precision BioSciences Announces In Vivo Gene Editing Collaboration with Novartis to Develop Potentially Curative Treatment for Disorders Including...