Genetic engineering

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Genetic engineering, also called genetic modification or genetic manipulation, is the modification and manipulation of an organism's

organisms. New DNA is obtained by either isolating and copying the genetic material of interest using recombinant DNA methods or by artificially synthesising the DNA. A construct is usually created and used to insert this DNA into the host organism. The first recombinant DNA molecule was made by Paul Berg in 1972 by combining DNA from the monkey virus SV40 with the lambda virus. As well as inserting genes, the process can be used to remove, or "knock out", genes. The new DNA can be inserted randomly, or targeted to a specific part of the genome.[1]

An organism that is generated through genetic engineering is considered to be genetically modified (GM) and the resulting entity is a

bacterium generated by Herbert Boyer and Stanley Cohen in 1973. Rudolf Jaenisch created the first GM animal when he inserted foreign DNA into a mouse in 1974. The first company to focus on genetic engineering, Genentech, was founded in 1976 and started the production of human proteins. Genetically engineered human insulin was produced in 1978 and insulin-producing bacteria were commercialised in 1982. Genetically modified food has been sold since 1994, with the release of the Flavr Savr tomato. The Flavr Savr was engineered to have a longer shelf life, but most current GM crops are modified to increase resistance to insects and herbicides. GloFish, the first GMO designed as a pet, was sold in the United States in December 2003. In 2016 salmon
modified with a growth hormone were sold.

Genetic engineering has been applied in numerous fields including research, medicine, industrial biotechnology and agriculture. In research, GMOs are used to study gene function and expression through loss of function, gain of function, tracking and expression experiments. By knocking out genes responsible for certain conditions it is possible to create animal model organisms of human diseases. As well as producing hormones, vaccines and other drugs, genetic engineering has the potential to cure genetic diseases through gene therapy.Chinese hamster ovary (CHO) cells are used in industrial genetic engineering. Additionally mRNA vaccines are made through genetic engineering to treat viruses such as COVID-19. The same techniques that are used to produce drugs can also have industrial applications such as producing enzymes for laundry detergent, cheeses and other products.

The rise of commercialised genetically modified crops has provided economic benefit to farmers in many different countries, but has also been the source of most of the controversy surrounding the technology. This has been present since its early use; the first field trials were destroyed by anti-GM activists. Although there is a scientific consensus that currently available food derived from GM crops poses no greater risk to human health than conventional food, critics consider GM food safety a leading concern. Gene flow, impact on non-target organisms, control of the food supply and intellectual property rights have also been raised as potential issues. These concerns have led to the development of a regulatory framework, which started in 1975. It has led to an international treaty, the Cartagena Protocol on Biosafety, that was adopted in 2000. Individual countries have developed their own regulatory systems regarding GMOs, with the most marked differences occurring between the US and Europe.

IUPAC definition

Genetic engineering: Process of inserting new genetic information into existing cells in order to modify a specific organism for the purpose of changing its characteristics.

Note: Adapted from ref.[2][3]

Overview

cisgenic
genetic modification

Genetic engineering is a process that alters the genetic structure of an organism by either removing or introducing DNA, or modifying existing genetic material in situ. Unlike traditional animal and plant breeding, which involves doing multiple crosses and then selecting for the organism with the desired phenotype, genetic engineering takes the gene directly from one organism and delivers it to the other. This is much faster, can be used to insert any genes from any organism (even ones from different domains) and prevents other undesirable genes from also being added.[4]

Genetic engineering could potentially fix severe genetic disorders in humans by replacing the defective gene with a functioning one.[5] It is an important tool in research that allows the function of specific genes to be studied.[6] Drugs, vaccines and other products have been harvested from organisms engineered to produce them.[7] Crops have been developed that aid food security by increasing yield, nutritional value and tolerance to environmental stresses.[8]

The DNA can be introduced directly into the

host organism or into a cell that is then fused or hybridised with the host.[9] This relies on recombinant nucleic acid techniques to form new combinations of heritable genetic material followed by the incorporation of that material either indirectly through a vector system or directly through micro-injection, macro-injection or micro-encapsulation
.

Genetic engineering does not normally include traditional breeding, in vitro fertilisation, induction of polyploidy, mutagenesis and cell fusion techniques that do not use recombinant nucleic acids or a genetically modified organism in the process.[9] However, some broad definitions of genetic engineering include selective breeding.[10] Cloning and stem cell research, although not considered genetic engineering,[11] are closely related and genetic engineering can be used within them.[12] Synthetic biology is an emerging discipline that takes genetic engineering a step further by introducing artificially synthesised material into an organism.[13]

Plants, animals or microorganisms that have been changed through genetic engineering are termed

synonymous with genetic engineering while within the United States of America and Canada genetic modification can also be used to refer to more conventional breeding methods.[17][18][19]

History

Humans have altered the genomes of species for thousands of years through

James Watson and Francis Crick showed that the DNA molecule has a double-helix structure – though the general concept of direct genetic manipulation was explored in rudimentary form in Stanley G. Weinbaum's 1936 science fiction story Proteus Island.[25][26]

In 1974 Rudolf Jaenisch created a genetically modified mouse, the first GM animal.

In 1972,

transgenic animal[30] These achievements led to concerns in the scientific community about potential risks from genetic engineering, which were first discussed in depth at the Asilomar Conference in 1975. One of the main recommendations from this meeting was that government oversight of recombinant DNA research should be established until the technology was deemed safe.[31][32]

In 1976 Genentech, the first genetic engineering company, was founded by Herbert Boyer and

U.S. Supreme Court in the Diamond v. Chakrabarty case ruled that genetically altered life could be patented.[34] The insulin produced by bacteria was approved for release by the Food and Drug Administration (FDA) in 1982.[35]

In 1983, a biotech company, Advanced Genetic Sciences (AGS) applied for U.S. government authorisation to perform field tests with the ice-minus strain of Pseudomonas syringae to protect crops from frost, but environmental groups and protestors delayed the field tests for four years with legal challenges.[36] In 1987, the ice-minus strain of P. syringae became the first genetically modified organism (GMO) to be released into the environment[37] when a strawberry field and a potato field in California were sprayed with it.[38] Both test fields were attacked by activist groups the night before the tests occurred: "The world's first trial site attracted the world's first field trasher".[37]

The first field trials of

Environmental Protection Agency, after having been approved by the FDA, making it the first pesticide producing crop to be approved in the US.[43] In 2009 11 transgenic crops were grown commercially in 25 countries, the largest of which by area grown were the US, Brazil, Argentina, India, Canada, China, Paraguay and South Africa.[44]

In 2010, scientists at the

synthetic genome and inserted it into an empty bacterial cell. The resulting bacterium, named Mycoplasma laboratorium, could replicate and produce proteins.[45][46] Four years later this was taken a step further when a bacterium was developed that replicated a plasmid containing a unique base pair, creating the first organism engineered to use an expanded genetic alphabet.[47][48] In 2012, Jennifer Doudna and Emmanuelle Charpentier collaborated to develop the CRISPR/Cas9 system,[49][50] a technique which can be used to easily and specifically alter the genome of almost any organism.[51]

Process

Polymerase chain reaction is a powerful tool used in molecular cloning.

Creating a GMO is a multi-step process. Genetic engineers must first choose what gene they wish to insert into the organism. This is driven by what the aim is for the resultant organism and is built on earlier research.

transcriptomics and genome sequencing has made it much easier to find suitable genes.[52] Luck also plays its part; the Roundup Ready gene was discovered after scientists noticed a bacterium thriving in the presence of the herbicide.[53]

Gene isolation and cloning

The next step is to isolate the candidate gene. The

single-celled organisms, which makes it suitable as a genetic engineering tool.[59]

Before the gene is inserted into the target organism it must be combined with other genetic elements. These include a

antibiotic resistance, so researchers can easily determine which cells have been successfully transformed. The gene can also be modified at this stage for better expression or effectiveness. These manipulations are carried out using recombinant DNA techniques, such as restriction digests, ligations and molecular cloning.[60]

Inserting DNA into the host genome

biolistics
to insert DNA into plant tissue.

There are a number of techniques used to insert genetic material into the host genome. Some bacteria can naturally

viral vectors.[61]

Plant genomes can be engineered by physical methods or by use of

biolistics, where particles of gold or tungsten are coated with DNA and then shot into young plant cells,[64] and electroporation
, which involves using an electric shock to make the cell membrane permeable to plasmid DNA.

As only a single cell is transformed with genetic material, the organism must be

Selectable markers are used to easily differentiate transformed from untransformed cells. These markers are usually present in the transgenic organism, although a number of strategies have been developed that can remove the selectable marker from the mature transgenic plant.[68]

A. tumefaciens
attaching itself to a carrot cell

Further testing using PCR,

RT-PCR, Western blot, immunofluorescence, ELISA and phenotypic analysis.[70]

The new genetic material can be inserted randomly within the host genome or targeted to a specific location. The technique of

zinc finger nucleases,[73][74] transcription activator-like effector nucleases (TALENs),[75][76] and the Cas9-guideRNA system (adapted from CRISPR).[77][78] TALEN and CRISPR are the two most commonly used and each has its own advantages.[79] TALENs have greater target specificity, while CRISPR is easier to design and more efficient.[79] In addition to enhancing gene targeting, engineered nucleases can be used to introduce mutations at endogenous genes that generate a gene knockout.[80][81]

Applications

Genetic engineering has applications in medicine, research, industry and agriculture and can be used on a wide range of plants, animals and microorganisms.

genes knocked out, increased susceptibility to disease, hormones for extra growth and the ability to express proteins in their milk.[86]

Medicine

Genetic engineering has many applications to medicine that include the manufacturing of drugs, creation of

Genetic engineering is also used to create animal models of human diseases. Genetically modified mice are the most common genetically engineered animal model.[91] They have been used to study and model cancer (the oncomouse), obesity, heart disease, diabetes, arthritis, substance abuse, anxiety, aging and Parkinson disease.[92] Potential cures can be tested against these mouse models.

Gene therapy is the

genetic engineering of humans, generally by replacing defective genes with effective ones. Clinical research using somatic gene therapy has been conducted with several diseases, including X-linked SCID,[93] chronic lymphocytic leukemia (CLL),[94][95] and Parkinson's disease.[96] In 2012, Alipogene tiparvovec became the first gene therapy treatment to be approved for clinical use.[97][98] In 2015 a virus was used to insert a healthy gene into the skin cells of a boy suffering from a rare skin disease, epidermolysis bullosa, in order to grow, and then graft healthy skin onto 80 percent of the boy's body which was affected by the illness.[99]

human embryos,[102][103] leading scientists of major world academies to call for a moratorium on inheritable human genome edits.[104] There are also concerns that the technology could be used not just for treatment, but for enhancement, modification or alteration of a human beings' appearance, adaptability, intelligence, character or behavior.[105] The distinction between cure and enhancement can also be difficult to establish.[106] In November 2018, He Jiankui announced that he had edited the genomes of two human embryos, to attempt to disable the CCR5 gene, which codes for a receptor that HIV uses to enter cells. The work was widely condemned as unethical, dangerous, and premature.[107] Currently, germline modification is banned in 40 countries. Scientists that do this type of research will often let embryos grow for a few days without allowing it to develop into a baby.[108]

Researchers are altering the genome of pigs to induce the growth of human organs, with the aim of increasing the success of pig to human organ transplantation.[109] Scientists are creating "gene drives", changing the genomes of mosquitoes to make them immune to malaria, and then looking to spread the genetically altered mosquitoes throughout the mosquito population in the hopes of eliminating the disease.[110]

Research

Knockout mice
Human cells in which some proteins are fused with green fluorescent protein to allow them to be visualised

Genetic engineering is an important tool for

natural scientists, with the creation of transgenic organisms one of the most important tools for analysis of gene function.[111] Genes and other genetic information from a wide range of organisms can be inserted into bacteria for storage and modification, creating genetically modified bacteria in the process. Bacteria are cheap, easy to grow, clonal, multiply quickly, relatively easy to transform and can be stored at -80 °C almost indefinitely. Once a gene is isolated it can be stored inside the bacteria providing an unlimited supply for research.[112]

Organisms are genetically engineered to discover the functions of certain genes. This could be the effect on the phenotype of the organism, where the gene is expressed or what other genes it interacts with. These experiments generally involve loss of function, gain of function, tracking and expression.

Industrial

Products of genetic engineering

Organisms can have their cells transformed with a gene coding for a useful protein, such as an enzyme, so that they will

human growth hormone, and vaccines, supplements such as tryptophan, aid in the production of food (chymosin in cheese making) and fuels.[118] Other applications with genetically engineered bacteria could involve making them perform tasks outside their natural cycle, such as making biofuels,[119] cleaning up oil spills, carbon and other toxic waste[120] and detecting arsenic in drinking water.[121] Certain genetically modified microbes can also be used in biomining and bioremediation, due to their ability to extract heavy metals from their environment and incorporate them into compounds that are more easily recoverable.[122]

In materials science, a genetically modified virus has been used in a research laboratory as a scaffold for assembling a more environmentally friendly lithium-ion battery.[123][124] Bacteria have also been engineered to function as sensors by expressing a fluorescent protein under certain environmental conditions.[125]

Agriculture

lesser cornstalk borer larvae (top image).[126]

One of the best-known and

genetically modified livestock to produce genetically modified food. Crops have been developed to increase production, increase tolerance to abiotic stresses, alter the composition of the food, or to produce novel products.[127]

The first crops to be released commercially on a large scale provided protection from insect pests or tolerance to

herbicides. Fungal and virus resistant crops have also been developed or are in development.[128][129] This makes the insect and weed management of crops easier and can indirectly increase crop yield.[130][131] GM crops that directly improve yield by accelerating growth or making the plant more hardy (by improving salt, cold or drought tolerance) are also under development.[132] In 2016 Salmon have been genetically modified with growth hormones to reach normal adult size much faster.[133]

GMOs have been developed that modify the quality of produce by increasing the nutritional value or providing more industrially useful qualities or quantities.[132] The Amflora potato produces a more industrially useful blend of starches. Soybeans and canola have been genetically modified to produce more healthy oils.[134][135] The first commercialised GM food was a tomato that had delayed ripening, increasing its shelf life.[136]

Plants and animals have been engineered to produce materials they do not normally make. Pharming uses crops and animals as bioreactors to produce vaccines, drug intermediates, or the drugs themselves; the useful product is purified from the harvest and then used in the standard pharmaceutical production process.[137] Cows and goats have been engineered to express drugs and other proteins in their milk, and in 2009 the FDA approved a drug produced in goat milk.[138][139]

Other applications

Genetic engineering has potential applications in conservation and natural area management. Gene transfer through viral vectors has been proposed as a means of controlling invasive species as well as vaccinating threatened fauna from disease.[140] Transgenic trees have been suggested as a way to confer resistance to pathogens in wild populations.[141] With the increasing risks of maladaptation in organisms as a result of climate change and other perturbations, facilitated adaptation through gene tweaking could be one solution to reducing extinction risks.[142] Applications of genetic engineering in conservation are thus far mostly theoretical and have yet to be put into practice.

Genetic engineering is also being used to create microbial art.[143] Some bacteria have been genetically engineered to create black and white photographs.[144] Novelty items such as lavender-colored carnations,[145] blue roses,[146] and glowing fish[147][148] have also been produced through genetic engineering.

Regulation

The regulation of genetic engineering concerns the approaches taken by governments to assess and manage the risks associated with the development and release of GMOs. The development of a regulatory framework began in 1975, at

Asilomar meeting recommended a set of voluntary guidelines regarding the use of recombinant technology.[31] As the technology improved the US established a committee at the Office of Science and Technology,[150] which assigned regulatory approval of GM food to the USDA, FDA and EPA.[151] The Cartagena Protocol on Biosafety, an international treaty that governs the transfer, handling, and use of GMOs,[152] was adopted on 29 January 2000.[153] One hundred and fifty-seven countries are members of the Protocol, and many use it as a reference point for their own regulations.[154]

The legal and regulatory status of GM foods varies by country, with some nations banning or restricting them, and others permitting them with widely differing degrees of regulation.

irradiated food, are considered "new food" and subject to extensive, case-by-case, science-based food evaluation by the European Food Safety Authority. The criteria for authorisation fall in four broad categories: "safety", "freedom of choice", "labelling", and "traceability".[162]
The level of regulation in other countries that cultivate GMOs lie in between Europe and the United States.

Regulatory agencies by geographical region
Region Regulators Notes
US USDA, FDA and EPA[151]
Europe European Food Safety Authority[162]
Canada Health Canada and the Canadian Food Inspection Agency[163][164] Regulated products with novel features regardless of method of origin[165][166]
Africa Common Market for Eastern and Southern Africa[167] Final decision lies with each individual country.[167]
China Office of Agricultural Genetic Engineering Biosafety Administration[168]
India Institutional Biosafety Committee, Review Committee on Genetic Manipulation and Genetic Engineering Approval Committee[169]
Argentina National Agricultural Biotechnology Advisory Committee (environmental impact), the National Service of Health and Agrifood Quality (food safety) and the National Agribusiness Direction (effect on trade)[170] Final decision made by the Secretariat of Agriculture, Livestock, Fishery and Food.[170]
Brazil National Biosafety Technical Commission (environmental and food safety) and the Council of Ministers (commercial and economical issues)[170]
Australia Office of the Gene Technology Regulator (oversees all GM products), Therapeutic Goods Administration (GM medicines) and Food Standards Australia New Zealand (GM food).[171][172] The individual state governments can then assess the impact of release on markets and trade and apply further legislation to control approved genetically modified products.[172]

One of the key issues concerning regulators is whether GM products should be labeled. The

feed which contains greater than 0.9% of approved GMOs must be labelled.[161]

Controversy

Critics have objected to the use of genetic engineering on several grounds, including ethical, ecological and economic concerns. Many of these concerns involve GM crops and whether food produced from them is safe and what impact growing them will have on the environment. These controversies have led to litigation, international trade disputes, and protests, and to restrictive regulation of commercial products in some countries.[179]

Accusations that scientists are "playing God" and other religious issues have been ascribed to the technology from the beginning.[180] Other ethical issues raised include the patenting of life,[181] the use of intellectual property rights,[182] the level of labeling on products,[183][184] control of the food supply[185] and the objectivity of the regulatory process.[186] Although doubts have been raised,[187] economically most studies have found growing GM crops to be beneficial to farmers.[188][189][190]

soil microbes,[192] and an increase in secondary and resistant insect pests.[193][194] Many of the environmental impacts regarding GM crops may take many years to be understood and are also evident in conventional agriculture practices.[192][195] With the commercialisation of genetically modified fish there are concerns over what the environmental consequences will be if they escape.[196]

There are three main concerns over the safety of genetically modified food: whether they may provoke an

allergic reaction; whether the genes could transfer from the food into human cells; and whether the genes not approved for human consumption could outcross to other crops.[197] There is a scientific consensus[198][199][200][201] that currently available food derived from GM crops poses no greater risk to human health than conventional food,[202][203][204][205][206] but that each GM food needs to be tested on a case-by-case basis before introduction.[207][208][209] Nonetheless, members of the public are less likely than scientists to perceive GM foods as safe.[210][211][212][213]

In popular culture

Genetic engineering features in many

Tleilaxu.[215] Few films have informed audiences about genetic engineering, with the exception of the 1978 The Boys from Brazil and the 1993 Jurassic Park, both of which make use of a lesson, a demonstration, and a clip of scientific film.[216][217] Genetic engineering methods are weakly represented in film; Michael Clark, writing for the Wellcome Trust, calls the portrayal of genetic engineering and biotechnology "seriously distorted"[217] in films such as The 6th Day. In Clark's view, the biotechnology is typically "given fantastic but visually arresting forms" while the science is either relegated to the background or fictionalised to suit a young audience.[217]

In the 2007 video game, BioShock, genetic engineering plays an important role in the central storyline and universe. The game takes place in the fictional underwater dystopia Rapture, in which its inhabitants possess genetic superhuman abilities after injecting themselves with "plasmids", a serum which grants such powers. Also in the city of Rapture are "Little Sisters", little girls who are generically engineered, as well as a side-plot in which a cabaret singer sells her foetus to genetic scientists who implant false memories into the newborn and genetically engineer it to grow into an adult.

See also

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  201. ^ "Statement by the AAAS Board of Directors on Labeling of Genetically Modified Foods" (PDF). American Association for the Advancement of Science. 20 October 2012. Retrieved 8 February 2016. The EU, for example, has invested more than €300 million in research on the biosafety of GMOs. Its recent report states: "The main conclusion to be drawn from the efforts of more than 130 research projects, covering a period of more than 25 years of research and involving more than 500 independent research groups, is that biotechnology, and in particular GMOs, are not per se more risky than e.g. conventional plant breeding technologies." The World Health Organization, the American Medical Association, the U.S. National Academy of Sciences, the British Royal Society, and every other respected organization that has examined the evidence has come to the same conclusion: consuming foods containing ingredients derived from GM crops is no riskier than consuming the same foods containing ingredients from crop plants modified by conventional plant improvement techniques.Pinholster G (25 October 2012). "AAAS Board of Directors: Legally Mandating GM Food Labels Could "Mislead and Falsely Alarm Consumers"". American Association for the Advancement of Science. Retrieved 8 February 2016.
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  208. ^ Some medical organizations, including the British Medical Association, advocate further caution based upon the precautionary principle:"Genetically modified foods and health: a second interim statement" (PDF). British Medical Association. March 2004. Archived (PDF) from the original on 22 March 2014. Retrieved 21 March 2016. In our view, the potential for GM foods to cause harmful health effects is very small and many of the concerns expressed apply with equal vigour to conventionally derived foods. However, safety concerns cannot, as yet, be dismissed completely on the basis of information currently available. When seeking to optimise the balance between benefits and risks, it is prudent to err on the side of caution and, above all, learn from accumulating knowledge and experience. Any new technology such as genetic modification must be examined for possible benefits and risks to human health and the environment. As with all novel foods, safety assessments in relation to GM foods must be made on a case-by-case basis. Members of the GM jury project were briefed on various aspects of genetic modification by a diverse group of acknowledged experts in the relevant subjects. The GM jury reached the conclusion that the sale of GM foods currently available should be halted and the moratorium on commercial growth of GM crops should be continued. These conclusions were based on the precautionary principle and lack of evidence of any benefit. The Jury expressed concern over the impact of GM crops on farming, the environment, food safety and other potential health effects. The Royal Society review (2002) concluded that the risks to human health associated with the use of specific viral DNA sequences in GM plants are negligible, and while calling for caution in the introduction of potential allergens into food crops, stressed the absence of evidence that commercially available GM foods cause clinical allergic manifestations. The BMA shares the view that there is no robust evidence to prove that GM foods are unsafe but we endorse the call for further research and surveillance to provide convincing evidence of safety and benefit.
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Further reading

External links