Category Archives: Donald Trump

About three dozen former US officials who served under President Donald Trump jumped on a conference call this week to figure out how to thwart Trump's efforts in the 2022 and 2024 elections, according to CNN.

Jake Tapper reported on the call, which was said to have taken place last Monday and involved high-profile Trump officials including John Kelly, Trump's former chief of staff. Also on the call, per CNN, were the former White House staffers Alyssa Farah Griffin and Anthony Scaramucci, the former Department of Homeland Security official Elizabeth Neumann, and Chris Krebs, the former director of the Cybersecurity and Infrastructure Security Agency.

CNN noted that the people associated with the call were known critics of Trump. Kelly has made several statements about Trump, saying the former president "doesn't know any history at all, even some of the basics on the US." He also said in August 2020 that informing Trump that the things he planned to do were illegal was like "French kissing a chainsaw."

Scaramucci, who held the role of White House communications director for less than two weeks, has broken with Trump and repeatedly feuded with the former president. Griffin, a former spokeswoman for Vice President Mike Pence who also served as Trump's communications director, was ridiculed by the president, who called her a "clown." She also commented in January that the Republican Party was "morally in disrepair" because of the GOP's failure to acknowledge the January 6 Capitol riot was a "big deal."

According to Tapper, Miles Taylor, a Trump official turned prominent Trump critic, helped lead the call and told CNN the participants were "overflowing with ideas" on how to stop Trump. Ideas, according to Tapper, included "shining a light" on the former president's financial backers and figuring out how to defeat Trump-endorsed candidates running in 2022 and 2024.

"We all agreed passionately that letters and statements don't mean anything," Taylor told CNN. "The two operative words are 'electoral effects.' How can we have tangible electoral effects against the extremist candidates that have been endorsed by Trump?"

Taylor is best known for anonymously writing a scathing 2018 New York Times op-ed article titled "I Am Part of the Resistance Inside the Trump Administration."

The Trump-era White House press secretary Stephanie Grisham was said to have missed the call because of a COVID-19 infection but told CNN she was still engaged with the group. Grisham signaled the group's plans earlier this month, saying on January 6 that the group planned to meet to talk about how they could "formally do some things to try and stop" Trump.

The group which has not released its name or published a list of members is among several GOP-linked organizations that have expressed opposition to Trump. In October, a Republican group called Republicans for Voting Rights put up several billboards across the US, including in Times Square, to remind the former president that he lost the 2020 election. The Lincoln Project, founded by current and former Republicans, also worked during the 2020 election to prevent Trump's reelection.

Trump has not yet announced a 2024 presidential run and said in November that he would "probably" wait until after the 2022 midterm elections to confirm his decision on a 2024 presidential bid.

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Dozens of former Trump staff had call to figure out how to thwart Trump in 2022, 2024 - Business Insider

William Barr, Donald Trumps second attorney general and perceived hatchet man until he split from the former president over his lies about election fraud, will publish his memoirs in March.

HarperCollins, the publisher of One Damn Thing After Another: Memoirs of an Attorney General, promised a vivid and forthright read on Barrs long career in law and conservative politics, in which he was first attorney general under George HW Bush.

Barr takes readers behind the scenes during seminal moments of the Bush administration in the 1990s, from the LA riots to Pan Am 103 and Iran Contra, the publisher said on Tuesday.

With the Trump administration, Barr faced an unrelenting barrage of issues, such as Russia-gate, the opioid epidemic, Chinese espionage, big tech, the Covid outbreak, civil unrest, the first impeachment, and the 2020 election fallout.

The publisher also said Barr would help readers understand how Bush and Trump viewed power and justice at critical junctures of their presidencies.

During the investigation of Russian election meddling and links between Trump and Moscow, Barr stoked rage among Democrats who accused him of interfering on behalf of the president.

His handling of Robert Muellers report also prompted protest from the special counsel himself.

Republicans and other observers defended Barr but the Massachusetts senator Elizabeth Warren, then a candidate for the Democratic nomination for president, called him a disgrace and not a credible head of federal law enforcement.

Barr was also present during key flash points of the Trump administration, for instance, walking at the presidents side in summer 2020 when he marched across Washington DCs Lafayette Square, which had been cleared of protesters against racism and police brutality, to stage a photo op at a historic church.

Barr split from Trump as the president refused to admit defeat by Joe Biden in the 2020 election. Angry scenes between the two men have been reported in other books, including bestsellers by Bob Woodward and Robert Costa of the Washington Post and Jon Karl of ABC News.

Barr stoked Trumps rage by telling the Associated Press he had not seen evidence of fraud on a scale that could have effected a different outcome in the election.

He was no longer in office during the culmination of Trumps concerted attempt to overturn his election defeat the deadly Capitol riot of January 6.

On 7 January 2021, Barr condemned Trump for orchestrating a mob to pressure Congress and said: The presidents conduct was a betrayal of his office and supporters.

The same day, the Guardian published an article examining the state of the Department of Justice after Barrs second stint in the chair.

Vanita Gupta, a former head of the civil rights division, said: The morale and the reputation of the department has been gutted because of undue political influence on the decisions of career staff.

The department needs to be rebuilt by new leadership committed at every turn to decisions made on the law and on the facts, and not on what the president wants.

On Tuesday, Sadie Gurman, a Wall Street Journal reporter, was among observers to note the provenance and irony of Barrs chosen title.

Current attorney general Merrick Garland, Gurman said, might appreciate that Barrs book title is actually an homage to his hero, Ed Levi, who, when asked to describe the job of attorney general famously replied, Its just one damn thing after another.

Ed Levi, a law professor and non-politician, was installed by Gerald Ford in 1975, after the Watergate scandal brought down Richard Nixon and his attorney general, John Mitchell who served time in prison.

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Donald Trumps former attorney general William Barr to publish his memoirs - The Guardian

For months, the Justice Department has provided little public indication of whether, or how seriously, it is investigating the role played by former President Donald J. Trump in the violent attack on the Capitol last Jan. 6.

But on Tuesday, for the first time, evidence emerged in court papers that prosecutors have posed questions to at least one Jan. 6 defendant that were focused on establishing an organized conspiracy involving Mr. Trump and his allies to disrupt the work of Congress.

The papers were filed by a defense lawyer in the case of Brandon Straka, a former hair stylist who founded a group called the Walk Away Foundation, which seeks to persuade Democratic voters to leave the party.

On the day before the Capitol was stormed, Mr. Straka spoke at a pro-Trump rally in Washington with prominent right-wing figures like the conspiracy theorist Alex Jones. Mr. Straka was also at the Capitol on Jan. 6.

In October, he pleaded guilty to charges of disorderly conduct, admitting, among other things, that he had urged a crowd outside the building to wrest a riot shield away from a police officer.

Last week, prosecutors filed a sentencing memo in his case, recommending that he serve four months of home detention. The memo noted that Mr. Straka had met with prosecutors earlier this month as part of his plea agreement and had been cooperative in answering their questions.

It remains unclear exactly what those questions were, but Mr. Strakas lawyer, Bilal Essayli, offered a broad description in his clients own sentencing memo, which was filed on Tuesday. In the memo, Mr. Essayli said that during Mr. Strakas interview with prosecutors, the government was focused on establishing an organized conspiracy between defendant, President Donald J. Trump, and allies of the former president to disrupt the joint session of Congress on Jan. 6.

Mr. Straka answered all questions truthfully and denied the existence of any such plot, Mr. Essaylis memo said.

Mr. Essayli did not respond to phone calls seeking comment about the interview. William Miller, a spokesman for the U.S. attorneys office in Washington, which is prosecuting the cases related to Jan. 6, also declined to comment.

In the past year, prosecutors have charged more than 700 people in connection with the storming of the Capitol, including members of far-right extremist groups like the Oath Keepers militia, whose leader was accused of sedition last week with 10 of his subordinates.

Mark Meadows. Mr. Trumps chief of staff, who initially provided the panel with a trove of documents that showed the extent of his rolein the efforts to overturn the election, is now refusing to cooperate. The House voted to recommend holding Mr. Meadows in criminal contempt of Congress.

Scott Perry and Jim Jordan. The Republican representatives of Pennsylvaniaand Ohioare among a group of G.O.P. congressmenwho were deeply involved in efforts to overturn the election. Both Mr. Perryand Mr. Jordanhaverefused to cooperatewith the panel.

Big Tech firms. The panel has criticized Alphabet, Meta, Reddit and Twitterfor allowing extremism to spread on their platforms and saying they have failed to cooperate adequately with the inquiry. The committee has issued subpoenas to all four companies.

Michael Flynn. Mr. Trumps former national security adviser attended an Oval Office meeting on Dec. 18 in which participants discussed seizing voting machines and invoking certain national security emergency powers. Mr. Flynn has filed a lawsuitto block the panels subpoenas.

Phil Waldron. The retired Army colonelhas been under scrutiny since a 38-page PowerPoint documenthe circulated on Capitol Hill was turned over to the panel by Mr. Meadows. The document contained extreme plans to overturn the election.

John Eastman. The lawyer has been the subject of intense scrutinysince writing a memothat laid out how Mr. Trump could stay in power. Mr. Eastman was present at a meeting of Trump allies at the Willard Hotelthat has becomea prime focus of the panel.

Still, there has been enormous public interest in whether investigators will ultimately reach beyond those who took part in the melee on Jan. 6 and seek to build a case against Mr. Trump and the circle of his allies who helped inspire the violence that day with baseless claims of election fraud.

On the first anniversary of the attack, Attorney General Merrick B. Garland said he was committed to holding all Jan. 6 perpetrators, at any level, accountable under law. But he did not name Mr. Trump, and there has been little public indication of any effort by the department to seek information from or about any of his close aides or allies.

The description of the interviews in Mr. Strakas memo, however brief, echoed comments made by some members of the House select committee investigating the events of Jan. 6, who have questioned whether Mr. Trump broke the law by obstructing Congresss duty to oversee the peaceful transition of power to Joseph R. Biden Jr.

Representative Liz Cheney, Republican of Wyoming, the vice chairwoman of the House committee, has been particularly pointed in suggesting that Mr. Trump, by failing to stop the violence at the Capitol on Jan. 6, may have violated federal law. The law Ms. Cheney was referring to obstruction of an official proceeding before Congress has been challenged repeatedly by defense lawyers with Jan. 6 cases, but five separate federal judges have recently ruled that it was viable.

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Prosecutors Reportedly Asked About Trumps Role in Jan. 6 Riot - The New York Times

Former President Donald Trump and his family company have a long history of stiffing contractors, but theres one bill they almost certainly wish they had paid.

Ahead of the 2017 presidential inauguration, the Trump Organization reserved a block of rooms at the Loews Madison Hotel. When at least 13 people didnt show up, the Trump Organization refused to pay the bill, something it has done many times in the past. The company then dodged a credit collection agency and eventually squirmed out of it by pushing the $49,358 bill off to the nonprofit presidential inaugural committee, the PIC.

That dodged payment is now the crux of the attorney general for the District of Columbias latest effort to put the Trump Organization back in its crosshairs in an ongoing investigation into how the Trump kids used the Presidential Inauguration Committee to throw lavish parties of their own.

It was their friends. It should never have been sent to the PIC. Thats misuse of funding. The Trump Organization being involved in any way and getting the PIC to pay any sort of balance anywhere on their behalf? It just doesnt seem legitimate, said Stephanie Winston Wolkoff, who coordinated inaugural events and is now the governments lead witness in this case.

Stephanie Winston Wolkoff

Drew Angerer/Getty

Winston Wolkoff is no friend of the Trumpsany more. Although she was close to the family for more than a decade and eventually became Trusted Adviser to First Lady Melania Trump, there was a fallout after Winston Wolkoff felt that the Trump White House made her the scapegoat for inauguration misspending. The New York Times identified a company associated with her, WIS Media Partners, as the recipient of a whopping $26 million, and Winston Wolkoff later overcame Justice Department resistance to publication of her tell-all book called Melania and Me.

D.C. Attorney General Karl Racine continues to investigate how the inauguration committee allegedly misspent more than $1 million and was allegedly used to essentially enrich Trumps own company on his way into the White House. And the Attorney Generals office is trying to recover from a courtroom defeat late last year.

In November, D.C. Superior Court Judge Jos M. Lpez seemed to doom the local attorney generals investigation when he cut the Trump Organization loose from the lawsuit. His reasoning, which surprised those following the case, was that the familys company wasnt directly involvedeven though Don Jr., Ivanka, and other staffers at the companys New York office were on a lot of the paperwork. So he dropped the Trump Organization from the lawsuit.

Washington, D.C., Attorney General Karl Racine

Alex Wong/Getty

The judges Nov. 8 order hinged on the companys claims that Texas financier Gentry Beach didnt have the authority to list the Trump Organization when he pulled out an American Express credit card and made the large and expensive reservation. However, Beach was no stranger to the Trump Organization. He was Donald Trump Jr.s college pal and was handpicked to serve on the nonprofits finance committee. He was also outed by journalists even before the inauguration for being part of a nonprofitdirected by Eric Trump, Don Jr. and another wealthy Texanthat seemed to be auctioning off access to the Trumps.

Since then, Racines office has filed documents in court seeking to reverse that, pointing to numerous receipts and memos that show how even the debt collector wouldnt be duped into letting the Trump Organization weasel its way out of this one.

In the typical fashion of an aggressive collections agency, Campbell Hightower & Adams in Arizona started bombarding the company with phone calls and emails in June 2017, picking up where the Loews Madison Hotel had left off.

A collector, identified only as Sherie, jotted down notes when she repeatedly communicated with Don Jr.s executive assistant, Kara Hanley.

Unfortunately, this was not an agreement made by anyone at The Trump Organization. Best, Kara, Hanley wrote on June 8.

A contract was signed and then 13 people did not show up for the rooms you reserved [sic] so according to the contract terms, those rooms still have to be paid for. What am I missing? Sherie wrote back.

A few weeks later, Sherie notified the Trump Organization that she had just found out that yet another Don Jr. executive assistant, Lindsey Santoro, had initially requested the rooms and added Beach as the main contact for the deal. That information seemed to cement even further that the company was indeed involved.

And when the hotel contacted the collections agency in July to request that the bill suddenly change the listed debtor to 58th Presidential Inauguration Committeewith an odd special note saying It just cannot say The Trump OrganizationSherie grew suspicious.

I hesitate as they all seem to be pointing fingers and making excuses as to why they wont pay it and this seems to be another ploy so the Trump Organizations name is not on it, Sherie wrote back on July 10.

Records show the bill was eventually paid by the Presidential Inaugural Committee at the direction of Rick Gates, a Trump political operator ally who served on the committeeand eventually served jail time for committing unrelated crimes caught by special counsel Robert Mueller's Russia investigation.

The District of Columbias AG hopes this evidence proves that the Trump Organization should remain part of the lawsuit, which seeks to seize money it deems was misused and divert it instead to another nonprofit. Otherwise, the civil investigation would continue only against the PIC (which is no longer active) and the Trump International Hotel Washington (which is being sold anyway).

In court filings, a lawyer for the Trump Organization and Trump Hotel blasted the AGs last-ditch effort as merely rehashed arguments that seek several bites at several apples. On Dec. 14, attorney Rebecca Woods reiterated that Don Jr.s buddy, Beach, didnt have the explicit authority to make the deal. She also wrote that investigators shouldnt be allowed to now seek sworn testimony from him or Santoro, the executive assistant.

When approached by The Daily Beast, the AGs office pointed to the arguments it made in court. The Trump Organizations lawyer didnt respond to a request for comment. The collection agency didnt return calls on Friday.

Notably, none of these documents described yet another layer of Trump Organization involvement: how company chief financial officer Allen Weisselberg puzzlingly assumed the responsibility of auditing the nonprofit PICs finances. Last summer, D.C. investigators wanted to interview him under oath, but he was then indicted for criminal tax fraud in New York City.

The local attorney generals request is now up to the judgebut a different one this time. On New Years Eve, the case was reassigned to D.C. Superior Court Judge Yvonne Williams, a former NAACP lawyer appointed to the bench by President Barack Obama.

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The Trump Org Stiffed a Hotel. His Kids May Pay the Price. - The Daily Beast

A look at some of the top headlines trending online today around the world including what you need to know about potential problems for Donald Trump, updates on the Russia-Ukraine standoff, the latest developments surrounding the Omicron surge and much more.

N.Y. Attorney General Outlines Pattern of Possible Fraud at Trump Business (NY Times)

Rudy Giuliani, three other Trump allies subpoenaed in January 6 riot probe (CNBC)

Former Trump officials plotting effort to blunt his impact on elections: report (The Hill)

Blinken launches fresh diplomatic push in Ukraine as fears of Russian invasion mount (NBC)

Airlines scrap or change flights to U.S. over 5G dispute, even though technologys rollouts been delayed near some airports (AP)

Federal N95 mask giveaway will start soon (cleveland.com)

Omicron is not that mild: 50,000 to 300,000 more US deaths projected by March (USA Today)

Nearly 1 million pediatric Covid cases reported last week (NBC)

COVID-19 health emergency could be over this year, WHO says (ABC)

Lawmakers call for investigation into at-home COVID test price gouging (CBS)

Fashion Icon and Former Vogue Creative Director Andr Leon Talley Dead at 73 (People)

Dallas Cowboys Dak Prescott apologizes for comments about fans throwing objects at officials (ESPN)

Harder They Fall, Insecure, H.E.R. nab NAACP awards noms (AP)

Microsoft buys gaming giant Activision Blizzard for $68.7 billion (AP)

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Omicron news, Donald Trump, Russia & more: Whats trending today - cleveland.com

Here are five things you must know for Wednesday, January 19:

U.S. equity futures edged higher Wednesday, while Treasury yields extended their recent surge amid bets that the Federal Reserve will quicken the pace of near-term rate hikes as inflationary pressures continue to build in the world's largest economy.

Stocks were hit hard by the prospects of faster rate hikes yesterday, while weaker-than-expected quarterly earnings from Goldman Sachs GS kept the Dow Jones Industrial Average deeply in the red, closing out its worst day since November, and pushed the S&P 500 into a year-to-date decline of around 4%.

A 0.25% March rate hike is all but assured from the Fed, but bets on a 50 basis point move are starting to creep in, lifting 2-year Treasury note yields to a February 2020 high of 1.075%.

Further upside pressures on oil prices, which have lifted WTI crude to the highest levels in seven years, were evident overnight amid disruption in a pipeline between Iraq and Turkey, while benchmark 10-year German bund yields traded in positive territory for the first time since 2019.

Bond market moves are likely to loom large over the markets again Wednesday, although a series of blue-chip earnings reports -- including updates from Dow components UnitedHealth (UNH) - Get UnitedHealth Group Incorporated Report and Procter & Gamble (PG) - Get Procter & Gamble Company Report-- will also provide direction prior to the start of trading.

Futures tied to the Dow are indicating a modest 35 point opening bell gain while those linked to the S&P 500 are priced for an 8 point bump to the upside.

Nasdaq Composite futures are indicating a 40 point opening bell gain as benchmark 10-year Treasury note yields climb to a post-pandemic high of 1.895% in overnight trading.

Ford F shares edged lower in pre-market trading after the carmaker said late Tuesday that its early investment in Rivian Automotive (RIVN) - Get Rivian Automotive, Inc. Class A Reportwould add around $8.2 billion to its fourth quarter bottom line.

In an investor update ahead of its formal earnings release on February 3, Ford said gains from Rivian's November IPO would be booked as a special item over the three months ending in December. Ford invested $500 million in Rivian in 2019 and has a total stake of around 12% in the Irvine, California-based EV group.

Rivian shares, once valued at more than $100 billion, closed at $73.16 last night, pegging their market cap at just under $66 billion.

Ford shares were marked 0.12% lower in pre-market trading Wednesday to indicate an opening bell price of $24.35 each.

U.S. air travelers could face another day of delay and disruption Wednesday as carriers around the world cancel flights in and out of the country following the initial rollout of 5G networks by AT&T (T) - Get AT&T Inc. Reportand Verizon (VZ) - Get Verizon Communications Inc. Report.

Although the wireless groups said they would pause rollouts near major U.S. airports, following warnings from both airline bosses and the Federal Aviation Administration linked to concerns that the 5G frequencies could affect some flight navigation instruments -- particularly in planes made by Boeing (BA) - Get Boeing Company Report-- several international carriers have cancelled flights or substituted aircraft to U.S. destinations.

President Joe Biden said the agreement to pause 5G rollouts at key airport towers would "avoid potentially devastating disruptions to passenger travel, cargo operations, and our economic recovery, while allowing more than 90% of wireless tower deployment to occur as scheduled."

United Airlines (UAL) - Get United Airlines Holdings, Inc. Reportwas marked 0.1% lower in pre-market trading at $45.60 each, while American Airlines (AAL) - Get American Airlines Group, Inc. Reportedged 0.2% higher to $17.94. Delta Air Lines (DAL) - Get Delta Air Lines, Inc. Reportwas marked 0.15% higher at $39.60 each while Boeing was little-changed at $225.05 each.

U.S. chip stocks jumped higher in pre-market trading aftersemiconductor equipment makerASML NV (AMSL) posted stronger-than-expected fourth quarter earnings and issued a robust near-term outlook for the sector.

ASML said demand for its extreme ultraviolet lithography systems, or EUV, machines, which design complex chips used by, sector titans such as Samsung Electronics, Intel and Taiwan Semiconductor and cost as much as $120 million, would help overall sales grow more than 20% this year, easing concerns that a fire in one of its German-based factories would impact supplies.

ASML's bullish outlook follows a similar assessment from TaiwanSemi (TSM) - Get Taiwan Semiconductor Manufacturing Co. Ltd. Reportthe world's biggest contract chipmaker and a lead supplier for Apple Inc. (AAPL) - Get Apple Inc. ReportiPhones, which posted record fourth quarter profits last week and boosted itscompound annual growth rate targets and capital spending plans.

Nvidia (NVDA) - Get NVIDIA Corporation Reportshares were marked 0.5% higher in pre-market trading Wednesday while Advanced Micro Devices (AMD) - Get Advanced Micro Devices, Inc. Reportgained 0.1% to $132.00 each. Intel (INTC) - Get Intel Corporation Reportwas marked 0.2% higher at $54.86 while Micron (MU) - Get Micron Technology, Inc. Report gained 0.85% to $93.65 each.

Former President Donald Trump was accused by the state of New York of using"fraudulent or misleading" asset valuations to obtain loans and tax deductions for his family organization.

New York Attorney General Letitia James said Trump, as well as his son Donald Jr. and daughter Ivanka, must provide sworn testimony as part of her investigation into the Trump Organization's financial affairs.

"Thus far in our investigation, we have uncovered significant evidence that suggests Donald J. Trump and the Trump Organization falsely and fraudulently valued multiple assets and misrepresented those values to financial institutions for economic benefit," James said in a statement.

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Stocks Edge Higher, Ford Cashes In, 5G Rollout Pause, Bullish ASML And Donald Trump - 5 Things You Must Know - TheStreet

Learning Objectives

By the end of this section, you will be able to:

Biotechnology is the use of artificial methods to modify the genetic material of living organisms or cells to produce novel compounds or to perform new functions. Biotechnology has been used for improving livestock and crops since the beginning of agriculture through selective breeding. Since the discovery of the structure of DNA in 1953, and particularly since the development of tools and methods to manipulate DNA in the 1970s, biotechnology has become synonymous with the manipulation of organisms DNA at the molecular level. The primary applications of this technology are in medicine (for the production of vaccines and antibiotics) and in agriculture (for the genetic modification of crops). Biotechnology also has many industrial applications, such as fermentation, the treatment of oil spills, and the production of biofuels, as well as many household applications such as the use of enzymes in laundry detergent.

To accomplish the applications described above, biotechnologists must be able to extract, manipulate, and analyze nucleic acids.

To understand the basic techniques used to work with nucleic acids, remember that nucleic acids are macromolecules made of nucleotides (a sugar, a phosphate, and a nitrogenous base). The phosphate groups on these molecules each have a net negative charge. An entire set of DNA molecules in the nucleus of eukaryotic organisms is called the genome. DNA has two complementary strands linked by hydrogen bonds between the paired bases.

Unlike DNA in eukaryotic cells, RNA molecules leave the nucleus. Messenger RNA (mRNA) is analyzed most frequently because it represents the protein-coding genes that are being expressed in the cell.

To study or manipulate nucleic acids, the DNA must first be extracted from cells. Various techniques are used to extract different types of DNA (Figure 10.2). Most nucleic acid extraction techniques involve steps to break open the cell, and then the use of enzymatic reactions to destroy all undesired macromolecules. Cells are broken open using a detergent solution containing buffering compounds. To prevent degradation and contamination, macromolecules such as proteins and RNA are inactivated using enzymes. The DNA is then brought out of solution using alcohol. The resulting DNA, because it is made up of long polymers, forms a gelatinous mass.

RNA is studied to understand gene expression patterns in cells. RNA is naturally very unstable because enzymes that break down RNA are commonly present in nature. Some are even secreted by our own skin and are very difficult to inactivate. Similar to DNA extraction, RNA extraction involves the use of various buffers and enzymes to inactivate other macromolecules and preserve only the RNA.

Because nucleic acids are negatively charged ions at neutral or alkaline pH in an aqueous environment, they can be moved by an electric field. Gel electrophoresis is a technique used to separate charged molecules on the basis of size and charge. The nucleic acids can be separated as whole chromosomes or as fragments. The nucleic acids are loaded into a slot at one end of a gel matrix, an electric current is applied, and negatively charged molecules are pulled toward the opposite end of the gel (the end with the positive electrode). Smaller molecules move through the pores in the gel faster than larger molecules; this difference in the rate of migration separates the fragments on the basis of size. The nucleic acids in a gel matrix are invisible until they are stained with a compound that allows them to be seen, such as a dye. Distinct fragments of nucleic acids appear as bands at specific distances from the top of the gel (the negative electrode end) that are based on their size (Figure 10.3). A mixture of many fragments of varying sizes appear as a long smear, whereas uncut genomic DNA is usually too large to run through the gel and forms a single large band at the top of the gel.

DNA analysis often requires focusing on one or more specific regions of the genome. It also frequently involves situations in which only one or a few copies of a DNA molecule are available for further analysis. These amounts are insufficient for most procedures, such as gel electrophoresis. Polymerase chain reaction (PCR) is a technique used to rapidly increase the number of copies of specific regions of DNA for further analyses (Figure 10.4). PCR uses a special form of DNA polymerase, the enzyme that replicates DNA, and other short nucleotide sequences called primers that base pair to a specific portion of the DNA being replicated. PCR is used for many purposes in laboratories. These include: 1) the identification of the owner of a DNA sample left at a crime scene; 2) paternity analysis; 3) the comparison of small amounts of ancient DNA with modern organisms; and 4) determining the sequence of nucleotides in a specific region.

In general, cloning means the creation of a perfect replica. Typically, the word is used to describe the creation of a genetically identical copy. In biology, the re-creation of a whole organism is referred to as reproductive cloning. Long before attempts were made to clone an entire organism, researchers learned how to copy short stretches of DNAa process that is referred to as molecular cloning.

Cloning allows for the creation of multiple copies of genes, expression of genes, and study of specific genes. To get the DNA fragment into a bacterial cell in a form that will be copied or expressed, the fragment is first inserted into a plasmid. A plasmid (also called a vector in this context) is a small circular DNA molecule that replicates independently of the chromosomal DNA in bacteria. In cloning, the plasmid molecules can be used to provide a vehicle in which to insert a desired DNA fragment. Modified plasmids are usually reintroduced into a bacterial host for replication. As the bacteria divide, they copy their own DNA (including the plasmids). The inserted DNA fragment is copied along with the rest of the bacterial DNA. In a bacterial cell, the fragment of DNA from the human genome (or another organism that is being studied) is referred to as foreign DNA to differentiate it from the DNA of the bacterium (the host DNA).

Plasmids occur naturally in bacterial populations (such as Escherichia coli) and have genes that can contribute favorable traits to the organism, such as antibiotic resistance (the ability to be unaffected by antibiotics). Plasmids have been highly engineered as vectors for molecular cloning and for the subsequent large-scale production of important molecules, such as insulin. A valuable characteristic of plasmid vectors is the ease with which a foreign DNA fragment can be introduced. These plasmid vectors contain many short DNA sequences that can be cut with different commonly available restriction enzymes. Restriction enzymes (also called restriction endonucleases) recognize specific DNA sequences and cut them in a predictable manner; they are naturally produced by bacteria as a defense mechanism against foreign DNA. Many restriction enzymes make staggered cuts in the two strands of DNA, such that the cut ends have a 2- to 4-nucleotide single-stranded overhang. The sequence that is recognized by the restriction enzyme is a four- to eight-nucleotide sequence that is a palindrome. Like with a word palindrome, this means the sequence reads the same forward and backward. In most cases, the sequence reads the same forward on one strand and backward on the complementary strand. When a staggered cut is made in a sequence like this, the overhangs are complementary (Figure 10.5).

Because these overhangs are capable of coming back together by hydrogen bonding with complementary overhangs on a piece of DNA cut with the same restriction enzyme, these are called sticky ends. The process of forming hydrogen bonds between complementary sequences on single strands to form double-stranded DNA is called annealing. Addition of an enzyme called DNA ligase, which takes part in DNA replication in cells, permanently joins the DNA fragments when the sticky ends come together. In this way, any DNA fragment can be spliced between the two ends of a plasmid DNA that has been cut with the same restriction enzyme (Figure 10.6).

Plasmids with foreign DNA inserted into them are called recombinant DNA molecules because they contain new combinations of genetic material. Proteins that are produced from recombinant DNA molecules are called recombinant proteins. Not all recombinant plasmids are capable of expressing genes. Plasmids may also be engineered to express proteins only when stimulated by certain environmental factors, so that scientists can control the expression of the recombinant proteins.

Reproductive cloning is a method used to make a clone or an identical copy of an entire multicellular organism. Most multicellular organisms undergo reproduction by sexual means, which involves the contribution of DNA from two individuals (parents), making it impossible to generate an identical copy or a clone of either parent. Recent advances in biotechnology have made it possible to reproductively clone mammals in the laboratory.

Natural sexual reproduction involves the union, during fertilization, of a sperm and an egg. Each of these gametes is haploid, meaning they contain one set of chromosomes in their nuclei. The resulting cell, or zygote, is then diploid and contains two sets of chromosomes. This cell divides mitotically to produce a multicellular organism. However, the union of just any two cells cannot produce a viable zygote; there are components in the cytoplasm of the egg cell that are essential for the early development of the embryo during its first few cell divisions. Without these provisions, there would be no subsequent development. Therefore, to produce a new individual, both a diploid genetic complement and an egg cytoplasm are required. The approach to producing an artificially cloned individual is to take the egg cell of one individual and to remove the haploid nucleus. Then a diploid nucleus from a body cell of a second individual, the donor, is put into the egg cell. The egg is then stimulated to divide so that development proceeds. This sounds simple, but in fact it takes many attempts before each of the steps is completed successfully.

The first cloned agricultural animal was Dolly, a sheep who was born in 1996. The success rate of reproductive cloning at the time was very low. Dolly lived for six years and died of a lung tumor (Figure 10.7). There was speculation that because the cell DNA that gave rise to Dolly came from an older individual, the age of the DNA may have affected her life expectancy. Since Dolly, several species of animals (such as horses, bulls, and goats) have been successfully cloned.

There have been attempts at producing cloned human embryos as sources of embryonic stem cells. In the procedure, the DNA from an adult human is introduced into a human egg cell, which is then stimulated to divide. The technology is similar to the technology that was used to produce Dolly, but the embryo is never implanted into a surrogate mother. The cells produced are called embryonic stem cells because they have the capacity to develop into many different kinds of cells, such as muscle or nerve cells. The stem cells could be used to research and ultimately provide therapeutic applications, such as replacing damaged tissues. The benefit of cloning in this instance is that the cells used to regenerate new tissues would be a perfect match to the donor of the original DNA. For example, a leukemia patient would not require a sibling with a tissue match for a bone-marrow transplant.

Why was Dolly a Finn-Dorset and not a Scottish Blackface sheep?

Because even though the original cell came from a Scottish Blackface sheep and the surrogate mother was a Scottish Blackface, the DNA came from a Finn-Dorset.

Using recombinant DNA technology to modify an organisms DNA to achieve desirable traits is called genetic engineering. Addition of foreign DNA in the form of recombinant DNA vectors that are generated by molecular cloning is the most common method of genetic engineering. An organism that receives the recombinant DNA is called a genetically modified organism (GMO). If the foreign DNA that is introduced comes from a different species, the host organism is called transgenic. Bacteria, plants, and animals have been genetically modified since the early 1970s for academic, medical, agricultural, and industrial purposes. These applications will be examined in more detail in the next module.

Watch this short video explaining how scientists create a transgenic animal.

Although the classic methods of studying the function of genes began with a given phenotype and determined the genetic basis of that phenotype, modern techniques allow researchers to start at the DNA sequence level and ask: What does this gene or DNA element do? This technique, called reverse genetics, has resulted in reversing the classical genetic methodology. One example of this method is analogous to damaging a body part to determine its function. An insect that loses a wing cannot fly, which means that the wings function is flight. The classic genetic method compares insects that cannot fly with insects that can fly, and observes that the non-flying insects have lost wings. Similarly in a reverse genetics approach, mutating or deleting genes provides researchers with clues about gene function. Alternately, reverse genetics can be used to cause a gene to overexpress itself to determine what phenotypic effects may occur.

Nucleic acids can be isolated from cells for the purposes of further analysis by breaking open the cells and enzymatically destroying all other major macromolecules. Fragmented or whole chromosomes can be separated on the basis of size by gel electrophoresis. Short stretches of DNA can be amplified by PCR. DNA can be cut (and subsequently re-spliced together) using restriction enzymes. The molecular and cellular techniques of biotechnology allow researchers to genetically engineer organisms, modifying them to achieve desirable traits.

Cloning may involve cloning small DNA fragments (molecular cloning), or cloning entire organisms (reproductive cloning). In molecular cloning with bacteria, a desired DNA fragment is inserted into a bacterial plasmid using restriction enzymes and the plasmid is taken up by a bacterium, which will then express the foreign DNA. Using other techniques, foreign genes can be inserted into eukaryotic organisms. In each case, the organisms are called transgenic organisms. In reproductive cloning, a donor nucleus is put into an enucleated egg cell, which is then stimulated to divide and develop into an organism.

In reverse genetics methods, a gene is mutated or removed in some way to identify its effect on the phenotype of the whole organism as a way to determine its function.

anneal: in molecular biology, the process by which two single strands of DNA hydrogen bond at complementary nucleotides to form a double-stranded molecule

biotechnology: the use of artificial methods to modify the genetic material of living organisms or cells to produce novel compounds or to perform new functions

cloning: the production of an exact copyspecifically, an exact genetic copyof a gene, cell, or organism

gel electrophoresis: a technique used to separate molecules on the basis of their ability to migrate through a semisolid gel in response to an electric current

genetic engineering: alteration of the genetic makeup of an organism using the molecular methods of biotechnologygenetically modified organism (GMO): an organism whose genome has been artificially changed

plasmid: a small circular molecule of DNA found in bacteria that replicates independently of the main bacterial chromosome; plasmids code for some important traits for bacteria and can be used as vectors to transport DNA into bacteria in genetic engineering applications

polymerase chain reaction (PCR): a technique used to make multiple copies of DNA

recombinant DNA: a combination of DNA fragments generated by molecular cloning that does not exist in naturestrong>recombinant protein: a protein that is expressed from recombinant DNA molecules

restriction enzyme: an enzyme that recognizes a specific nucleotide sequence in DNA and cuts the DNA double strand at that recognition site, often with a staggered cut leaving short single strands or sticky ends

reverse genetics: a form of genetic analysis that manipulates DNA to disrupt or affect the product of a gene to analyze the genes function

reproductive cloning: cloning of entire organisms

transgenic: describing an organism that receives DNA from a different species

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10.1 Cloning and Genetic Engineering Concepts of Biology ...

Despite all that controversy surrounding it, genetic engineering is here to stay and progress, as biomedical engineering technologies become smarter. Read about the different types of genetic engineering in the following article.

The advance of genetic engineering makes it quite conceivable that we will begin to design our own evolutionary progress.~ Isaac Asimov

Well, whether or not we begin to actually design our evolutionary progress is a different matter altogether but the fact that genetic engineering widens the scope of treating and curing various hereditary and terminal illnesses surely deserves some amount of attention and respect. You see, contrary to what most of us have been made to believe, thanks to those extravagant Hollywood sci-fi flicks, genetic engineering is not just about creating weird new species or making mutants out of men.

Genetic engineering, as it exists today, is more about improving the quality of the existing species of various organisms by way of enhancing their health, yield (in agriculture and livestock) and overall quality. This is usually done by tampering with the subjects genetic matter in such a way that their anatomies get conditioned to work in the survival mode, rather than the victim mode, in the face of disabling defects and life-threatening illnesses. For achieving these and similarly related ends, there exist three different types of genetic engineering, based upon their functions and fields of application. Lets proceed to the next segment and check out what they are.

The scope of genetic engineering is not restricted to curious human tampering of genetic paraphernalia in a bid to come up with various medical and scientific solutions. The greatest genetic engineer of all, Mother Nature, has been carrying out genetic manipulations all this time, since way long before the primate ancestors of humans were even introduced on Earth as a distinct species! Dont get me? Well, how else would you explain the phenomena of evolution, natural selection and selective breeding? Designing the blueprints of different versions of Software Life and creating appropriate hardware for each software version is no mean task! As far as human interest and efforts in genetic engineering are concerned, we are still way behind Nature and are way below the nascent stage. However, following are the three main categories of genetic engineering. Take a look.

This is the research branch of genetic engineering in which virtual genetic models are created using computer software. Various computer programs are used to theoretically study the implications of various genetic engineering activities if they are to be carried out in practice. For instance, before going ahead and splicing two different genes in actual practice, preparing an analytical model based upon an appropriate program, developed for the purpose, will give the researchers an idea whether such splicing would be successful at all and if successful, if the desired end would be achieved. This is a better way of carrying out the trial-and-error stage and reduces risks of disaster during experiments using real organisms, especially animals.

Applied genetic engineering, as the name suggests, is that field of genetic engineering which pertains to practical application of genetic engineering tools to manipulate the genes of living organisms for making genetic copies of them or to introduce certain different characteristics in them that are not usual for the subjects. The first instance is what we typically refer to as cloning and the second instance refers to the premises of transgenesis. While cloning is a highly regulated and controversial field, it has been carried out in various subjects of animal and plant species with mixed results and uncertain success rates. Transgenesis, on the other hand, is a comparatively common area and most of us have partaken of the results of transgenesis sometime or the other. Dont believe me? Well, what about hybrid fruits and vegetables? They are the most common and abundant examples of transgenesis.

Chemical genetic engineering can be called the grass root level of applied genetic engineering as it deals with separating, classifying and graphing genes to prepare them for applied genetic engineering activities and experiments. Chemical genetic engineering includes genetic mapping, studying genetic interaction and genetic coding. In genetic mapping, DNA fragments are assigned to individual chromosomes and thus, a genetic map is created after the complete DNA sequencing of a subject is done. Genetic mapping is very crucial to understanding the disease-gene link and this understanding lays the foundation of various gene therapies. Studying genetic interactions helps researchers understand exactly what set and combination of genes would produce a particular phenotype or set of morphological, physiological and behavioral characteristics. Genetic coding deals with studying and experimenting with amino acid sequences of DNA and RNA so as to understand the heredity trends and characteristics of a subject. This helps in understanding the bases, possibilities and conditions of undesirable hereditary characteristics, defects and disease in a bid to come out with medical solutions for the same.

So, those were the three major categories of genetic engineering that have been developed till date. Other than the aforementioned genetic engineering types, two emerging fields in genetic engineering are somatic cell engineering and germ-line engineering. Both of these are, at present, the most significant types of genetic engineering in humans and both are used in gene therapy for correcting defective genes and for preventing the transmission of hereditary defects or diseases from one generation to subsequent generations. Both of these classes of genetic engineering hold the potential of coming up with effective cures for degenerative conditions such as Parkinsons, Alzheimers and Huntingtons disease.

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Types of Genetic Engineering - Biology Wise

Abstract

Genetic engineering opens new possibilities for biomedical enhancement requiring ethical, societal and practical considerations to evaluate its implications for human biology, human evolution and our natural environment. In this Commentary, we consider human enhancement, and in particular, we explore genetic enhancement in an evolutionary context. In summarizing key open questions, we highlight the importance of acknowledging multiple effects (pleiotropy) and complex epigenetic interactions among genotype, phenotype and ecology, and the need to consider the unit of impact not only to the human body but also to human populations and their natural environment (systems biology). We also propose that a practicable distinction between therapy and enhancement may need to be drawn and effectively implemented in future regulations. Overall, we suggest that it is essential for ethical, philosophical and policy discussions on human enhancement to consider the empirical evidence provided by evolutionary biology, developmental biology and other disciplines.

Lay Summary: This Commentary explores genetic enhancement in an evolutionary context. We highlight the multiple effects associated with germline heritable genetic intervention, the need to consider the unit of impact to human populations and their natural environment, and propose that a practicable distinction between therapy and enhancement is needed.

There are countless examples where technology has contributed to ameliorate the lives of people by improving their inherent or acquired capabilities. For example, over time, there have been biomedical interventions attempting to restore functions that are deficient, such as vision, hearing or mobility. If we consider human vision, substantial advances started from the time spectacles were developed (possibly in the 13th century), continuing in the last few years, with researchers implanting artificial retinas to give blind patients partial sight [13]. Recently, scientists have also successfully linked the brain of a paralysed man to a computer chip, which helped restore partial movement of limbs previously non-responsive [4, 5]. In addition, synthetic blood substitutes have been created, which could be used in human patients in the future [68].

The progress being made by technology in a restorative and therapeutic context could in theory be applied in other contexts to treat non-pathological conditions. Many of the technologies and pharmaceutical products developed in a medical context to treat patients are already being used by humans to enhance some aspect of their bodies, for example drugs to boost brain power, nutritional supplements, brain stimulating technologies to control mood or growth hormones for children of short stature. Assistive technology for disabled people, reproductive medicine and pharmacology, beside their therapeutic and restorative use, have a greater potential for human enhancement than currently thought. There are also dual outcomes as some therapies can have effects that amount to an enhancement as for example, the artificial legs used by the South African sprinter Oscar Pistorius providing him with a competitive advantage.

This commentary will provide general ethical considerations on human enhancement, and within the several forms of so-called human biomedical enhancement, it will focus on genetic engineering, particularly on germline (heritable) genetic interventions and on the insights evolutionary biology can provide in rationalizing its likely impact. These insights are a subject often limited in discussions on genetic engineering and human enhancement in general, and its links to ethical, philosophical and policy discussions, in particular [9]. The rapid advances in genetic technology make this debate very topical. Moreover, genes are thought to play a very substantial role in biological evolution and development of the human species, thus making this a topic requiring due consideration. With this commentary, we explore how concepts based in evolutionary biology could contribute to better assess the implications of human germline modifications, assuming they were widely employed. We conclude our brief analysis by summarizing key issues requiring resolution and potential approaches to progress them. Overall, the aim is to contribute to the debate on human genetic enhancement by looking not only at the future, as it is so often done, but also at our evolutionary past.

The noun enhancement comes from the verb enhance, meaning to increase or improve. The verb enhance can be traced back to the vulgar Latin inaltiare and late Latin inaltare (raise, exalt), from altare (make high) and altus (high), literally grown tall. For centuries human enhancement has populated our imagination outlined by stories ranging from the myths of supernormal strengths and eternal life to the superpowers illustrated by the 20th century comic books superheroes. The desire of overcoming normal human capacities and the transformation to an almost perfect form has been part of the history of civilization, extending from arts and religion to philosophy. The goal of improving the human condition and health has always been a driver for innovation and biomedical developments.

In the broadest sense, the process of human enhancement can be considered as an improvement of the limitations of a natural version of the human species with respect to a specific reference in time, and to different environments, which can vary depending on factors such as, for example, climate change. The limitations of the human condition can be physical and/or mental/cognitive (e.g. vision, strength or memory). This poses relevant questions of what a real or perceived human limitation is in the environment and times in which we are living and how it can be shifted over time considering social norms and cultural values of modern societies. Besides, the impact that overcoming these limitations will have on us humans, and the environment, should also be considered. For example, if we boost the immune system of specific people, this may contribute to the development/evolution of more resistant viruses and bacteria or/and lead to new viruses and bacteria to emerge. In environmental terms, enhancing the longevity of humans could contribute to a massive increase in global population, creating additional pressures on ecosystems already under human pressure.

Two decades ago, the practices of human enhancement have been described as biomedical interventions that are used to improve human form or functioning beyond what is necessary to restore or sustain health [10]. The range of these practices has now increased with technological development, and they are any kind of genetic, biomedical, or pharmaceutical intervention aimed at improving human dispositions, capacities, or well-being, even if there is no pathology to be treated [11]. Practices of human enhancement could be visualized as upgrading a system, where interventions take place for a better performance of the original system. This is far from being a hypothetical situation. The rapid progress within the fields of nanotechnology, biotechnology, information technology and cognitive science has brought back discussions about the evolutionary trajectory of the human species by the promise of new applications which could provide abilities beyond current ones [12, 13]. If such a possibility was consciously embraced and actively pursued, technology could be expected to have a revolutionary interference with human life, not just helping humans in achieving general health and capabilities commensurate with our current ones but helping to overcome human limitations far beyond of what is currently possible for human beings. The emergence of new technologies has provided a broader range of potential human interventions and the possibility of transitioning from external changes to our bodies (e.g. external prosthesis) to internal ones, especially when considering genetic manipulation, whose changes can be permanent and transmissible.

The advocates of a far-reaching human enhancement have been referred to as transhumanists. In their vision, so far, humans have largely worked to control and shape their exterior environments (niche construction) but with new technologies (e.g. biotechnology, information technology and nanotechnology) they will soon be able to control and fundamentally change their own bodies. Supporters of these technologies agree with the possibility of a more radical interference in human life by using technology to overcome human limitations [1416], that could allow us to live longer, healthier and even happier lives [17]. On the other side, and against this position, are the so-called bioconservatives, arguing for the conservation and protection of some kind of human essence, with the argument that it exists something intrinsically valuable in human life that should be preserved [18, 19].

There is an ongoing debate between transhumanists [2022] and bioconservatives [18, 19, 23] on the ethical issues regarding the use of technologies in humans. The focus of this commentary is not centred on this debate, particularly because the discussion of these extreme, divergent positions is already very prominent in the public debate. In fact, it is interesting to notice that the moderate discourses around this topic are much less known. In a more moderate view, perhaps one of the crucial questions to consider, independently of the moral views on human enhancement, is whether human enhancement (especially if considering germline heritable genetic interventions) is a necessary development, and represents an appropriate use of time, funding and resources compared to other pressing societal issues. It is crucial to build space for these more moderate, and perhaps less polarized voices, allowing the consideration of other positions and visions beyond those being more strongly projected so far.

Ethical and societal discussions on what constitutes human enhancement will be fundamental to support the development of policy frameworks and regulations on new technological developments. When considering the ethical implications of human enhancement that technology will be available to offer now and in the future, it could be useful to group the different kinds of human enhancements in the phenotypic and genetic categories: (i) strictly phenotypic intervention (e.g. ranging from infrared vision spectacles to exoskeletons and bionic limbs); (ii) somatic, non-heritable genetic intervention (e.g. editing of muscle cells for stronger muscles) and (iii) germline, heritable genetic intervention (e.g. editing of the CC chemokine receptor type 5 (CCR5) gene in the Chinese baby twins, discussed later on). These categories of enhancement raise different considerations and concerns and currently present different levels of acceptance by our society. The degree of ethical, societal and environmental impacts is likely to be more limited for phenotypic interventions (i) but higher for genetic interventions (ii and iii), especially for the ones which are transmissible to future generations (iii).

The rapid advances in technology seen in the last decades, have raised the possibility of radical enhancement, defined by Nicholas Agar, as the improvement of human attributes and abilities to levels that greatly exceed what is currently possible for human beings [24]. Genetic engineering offers the possibility of such an enhancement by providing humans a profound control over their own biology. Among other technologies, genetic engineering comprises genome editing (also called gene editing), a group of technologies with the ability to directly modify an organisms DNA through a targeted intervention in the genome (e.g. insertion, deletion or replacement of specific genetic material) [25]. Genome editing is considered to achieve much greater precision than pre-existing forms of genetic engineering. It has been argued to be a revolutionary tool due to its efficiency, reducing cost and time. This technology is considered to have many applications for human health, in both preventing and tackling disease. Much of the ethical debate associated with this technology concerns the possible application of genome editing in the human germline, i.e. the genome that can be transmitted to following generations, be it from gametes, a fertilized egg or from first embryo divisions [2628]. There has been concern as well as enthusiasm on the potential of the technology to modify human germline genome to provide us with traits considered positive or useful (e.g. muscle strength, memory and intelligence) in the current and future environments.

To explore some of the possible implications of heritable interventions we will take as an example the editing (more specifically deletion using CRISPR genome editing technology) of several base pairs of the CCR5 gene. Such intervention was practised in 2018 in two non-identical twin girls born in China. Loss of function mutations of the CCR5 had been previously shown to provide resistance to HIV. Therefore, the gene deletion would be expected to protect the twin baby girls from risk of transmission of HIV which could have occurred from their father (HIV-positive). However, the father had the infection kept under control and the titre of HIV virus was undetectable, which means that risk of transmission of HIV infection to the babies was negligible [29].

From an ethical ground, based on current acceptable practices, this case has been widely criticized by the scientific community beside being considered by many a case of human enhancement intervention rather than therapy [29, 30]. One of the questions this example helps illustrate is that the ethical boundary between a therapy that corrects a disorder by restoring performance to a normal scope, and an intervention that enhances human ability outside the accepted normal scope, is not always easy to draw. For the sake of argument, it could be assumed that therapy involves attempts to restore a certain condition of health, normality or sanity of the natural condition of a specific individual. If we take this approach, the question is how health, normality and sanity, as well as natural per se, are defined, as the meaning of these concepts shift over time to accommodate social norms and cultural values of modern societies. It could be said that the difficulty of developing a conceptual distinction between therapy and enhancement has always been present. However, the potential significance of such distinction is only now, with the acceleration and impact of technological developments, becoming more evident.

Beyond ethical questions, a major problem of this intervention is that we do not (yet?) know exactly the totality of the effects that the artificial mutation of the CCR5 may have, at both the genetic and phenotypic levels. This is because we now know that, contrary to the idea of one gene-one trait accepted some decades ago, a geneor its absencecan affect numerous traits, many of them being apparently unrelated (a phenomenon also known as pleiotropy). That is, due to constrained developmental interactions, mechanisms and genetic networks, a change in a single gene can result in a cascade of multiple effects [31]. In the case of CCR5, we currently know that the mutation offers protection against HIV infection, and also seems to increase the risk of severe or fatal reactions to some infectious diseases, such as the influenza virus [32]. It has also been observed that among people with multiple sclerosis, the ones with CCR5 mutation are twice as likely to die early than are people without the mutation [33]. Some studies have also shown that defective CCR5 can have a positive effect in cognition to enhance learning and memory in mice [34]. However, its not clear if this effect would be translated into humans. The example serves to illustrate that, even if human enhancement with gene editing methods was considered ethically sound, assessing the totality of its implications on solid grounds may be difficult to achieve.

Beyond providing the opportunity of enhancing human capabilities in specific individuals, intervening in the germline is likely to have an impact on the evolutionary processes of the human species raising questions on the scale and type of impacts. In fact, the use of large-scale genetic engineering might exponentially increase the force of niche construction in human evolution, and therefore raise ethical and practical questions never faced by our species before. It has been argued that natural selection is a mechanism of lesser importance in the case of current human evolution, as compared to other organisms, because of advances in medicine and healthcare [35]. According to such a view, among many others advances, natural selection has been conditioned by our niche-construction ability to improve healthcare and access to clean water and food, thus changing the landscape of pressures that humans have been facing for survival. An underlying assumption or position of the current debate is that, within our human species, the force of natural selection became minimized and that we are somehow at the end-point of our evolution [36]. If this premise holds true, one could argue that evolution is no longer a force in human history and hence that any human enhancement would not be substituting itself to human evolution as a key driver for future changes.

However, it is useful to remember that, as defined by Darwin in his book On the Origin of the Species, natural selection is a process in which organisms that happen to be better adapted to a certain environment tend to have higher survival and/or reproductive rates than other organisms [37]. When comparing human evolution to human genetic enhancement, an acceptable position could be to consider ethically sound those interventions that could be replicated naturally by evolution, as in the case of the CCR5 gene. Even if this approach was taken, however, it is important to bear in mind that human evolution acts on human traits sometimes increasing and sometimes decreasing our biological fitness, in a constant evolutionary trade-off and in a contingent and/or neutralin the sense of not progressiveprocess. In other worlds, differently from genetic human enhancement, natural selection does not aim at improving human traits [38]. Human evolution and the so-called genetic human enhancement would seem therefore to involve different underlying processes, raising several questions regarding the implications and risks of the latter.

But using genetic engineering to treat humans has been proposed far beyond the therapeutic case or to introduce genetic modifications known to already occur in nature. In particular, when looking into the views expressed on the balance between human evolution and genetic engineering, some argue that it may be appropriate to use genetic interventions to go beyond what natural selection has contributed to our species when it comes to eradicate vulnerabilities [17]. Furthermore, when considering the environmental, ecological and social issues of contemporary times, some suggest that genetic technologies could be crucial tools to contribute to human survival and well-being [2022]. The possible need to engineer human traits to ensure our survival could include the ability to allow our species to adapt rapidly to the rate of environmental change caused by human activity, for which Darwinian evolution may be too slow [39]. Or, for instance, to support long-distance space travel by engineering resistance to radiation and osteoporosis, along with other conditions which would be highly advantageous in space [40].

When considering the ethical and societal merits of these propositions, it is useful to consider how proto-forms of enhancement has been approached by past human societies. In particular, it can be argued that humans have already employedas part of our domestication/selective breeding of other animalstechniques of indirect manipulation of genomes on a relatively large scale over many millennia, albeit not on humans. The large-scale selective breeding of plants and animals over prehistoric and historic periods could be claimed to have already shaped some of our natural environment. Selective breeding has been used to obtain specific characteristics considered useful at a given time in plants and animals. Therefore, their evolutionary processes have been altered with the aim to produce lineages with advantageous traits, which contributed to the evolution of different domesticated species. However, differently from genetic engineering, domestication possesses inherent limitations in its ability to produce major transformations in the created lineages, in contrast with the many open possibilities provided by genetic engineering.

When considering the impact of genetic engineering on human evolution, one of questions to be considered concerns the effects, if any, that genetic technology could have on the genetic pool of the human population and any implication on its resilience to unforeseen circumstances. This underlines a relevant question associated with the difference between health and biological fitness. For example, a certain group of animals can be more healthyas domesticated dogsbut be less biologically fit according to Darwins definition. Specifically, if such group of animals are less genetically diverse than their ancestors, they could be less adaptable to environmental changes. Assuming that, the human germline modification is undertaken at a global scale, this could be expected to have an effect, on the distribution of genetically heritable traits on the human population over time. Considering that gene and trait distributions have been changing under the processes of evolution for billions of years, the impact on evolution will need to be assessed by analysing which genetic alterations have been eventually associated with specific changes within the recent evolutionary history of humans. On this front, a key study has analysed the implications of genetic engineering on the evolutionary biology of human populations, including the possibility of reducing human genetic diversity, for instance creating a biological monoculture [41]. The study argued that genetic engineering will have an insignificant impact on human diversity, while it would likely safeguard the capacity of human populations to deal with disease and new environmental challenges and therefore, ensure the health and longevity of our species [41]. If the findings of this study were considered consistent with other knowledge and encompassing, the impact of human genetic enhancements on the human genetic pool and associated impacts could be considered secondary aspects. However, data available from studies on domestication strongly suggests that domestication of both animals and plans might lead to not only decreased genetic diversity per se, but even affect patterns of variation in gene expression throughout the genome and generally decreased gene expression diversity across species [4244]. Given that, according to recent studies within the field of biological anthropology recent human evolution has been in fact a process of self-domestication [45], one could argue that studies on domestication could contribute to understanding the impacts of genetic engineering.

Beyond such considerations, it is useful to reflect on the fact that human genetic enhancement could occur on different geographical scales, regardless of the specific environment and geological periods in which humans are living and much more rapidly than in the case of evolution, in which changes are very slow. If this was to occur routinely and on a large scale, the implications of the resulting radical and abrupt changes may be difficult to predict and its impacts difficult to manage. This is currently highlighted by results of epigenetics studies, and also of the microbiome and of the effects of pollutants in the environment and their cumulative effect on the development of human and non-human organisms alike. Increasingly new evidence indicates a greater interdependence between humans and their environments (including other microorganisms), indicating that modifying the environment can have direct and unpredictable consequences on humans as well. This highlight the need of a systems level approach. An approach in which the bounded body of the individual human as a basic unit of biological or social action would need to be questioned in favour of a more encompassing and holistic unit. In fact, within biology, there is a new field, Systems Biology, which stresses the need to understand the role that pleiotropy, and thus networks at multiple levelse.g. genetic, cellular, among individuals and among different taxaplay within biological systems and their evolution [46]. Currently, much still needs to be understood about gene function, its role in human biological systems and the interaction between genes and external factors such as environment, diet and so on. In the future if we do choose to genetically enhance human traits to levels unlikely to be achieved by human evolution, it would be crucial to consider if and how our understanding of human evolution enable us to better understand the implications of genetic interventions.

New forms of human enhancement are increasingly coming to play due to technological development. If phenotypic and somatic interventions for human enhancement pose already significant ethical and societal challenges, germline heritable genetic intervention, require much broader and complex considerations at the level of the individual, society and human species as a whole. Germline interventions associated with modern technologies are capable of much more rapid, large-scale impacts and seem capable of radically altering the balance of humans with the environment. We know now that beside the role genes play on biological evolution and development, genetic interventions can induce multiple effects (pleiotropy) and complex epigenetics interactions among genotype, phenotype and ecology of a certain environment. As a result of the rapidity and scale with which such impact could be realized, it is essential for ethical and societal debates, as well as underlying scientific studies, to consider the unit of impact not only to the human body but also to human populations and their natural environment (systems biology). An important practicable distinction between therapy and enhancement may need to be drawn and effectively implemented in future regulations, although a distinct line between the two may be difficult to draw.

In the future if we do choose to genetically enhance human traits to levels unlikely to be achieved by human evolution, it would be crucial to consider if and how our understanding of humans and other organisms, including domesticated ones, enable us to better understand the implications of genetic interventions. In particular, effective regulation of genetic engineering may need to be based on a deep knowledge of the exact links between phenotype and genotype, as well the interaction of the human species with the environment and vice versa.

For a broader and consistent debate, it will be essential for technological, philosophical, ethical and policy discussions on human enhancement to consider the empirical evidence provided by evolutionary biology, developmental biology and other disciplines.

This work was supported by Fundao para a Cincia e a Tecnologia (FCT) of Portugal [CFCUL/FIL/00678/2019 to M.A.].

Conflict of interest: None declared.

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Human enhancement: Genetic engineering and evolution

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7 Future Genetic-Engineering Technologies | Genetically ...