Recombinant DNA

Definition

Recombinant DNA is a form of DNA constructed in the laboratory. It is generated by transferring selected pieces of DNA from one organism to another.

The vial shown in the photograph contains human insulin, one of the first therapeutic proteins that was genetically cloned. The drug is used to treat diabetes. Credit: Wellcome Library, London.

Importance

Genetic engineering is used for many different purposes in research, medicine, agriculture and industry. The technology is important because it enables the creation of multiple copies of genes and the insertion of foreign genes into other organisms to give them new traits, such as antibiotic resistance or a new colour. One of the first ways in which the technology was deployed was to re-engineer microbial cells to produce foreign proteins. This facilitated the manufacture of human proteins on an unprecedented scale at minimum cost, thereby opening the way to study the function of proteins in greater detail and to their therapeutic use. By 2001 over 80 recombinant DNA based products had been approved for treating disease and for vaccination and a further 350 recombinant DNA-based drugs were being tested for safety and efficacy. The technology is also an important tool in agriculture, being used to improve plants' resistance to pests and increase crop yields.

Discovery

While the structure of DNA was first determined in 1953, it was to take another two decades before scientists had the means to generate recombinant DNA. This was aided by firstly the realisation in the 1950s that plasmids, small mobile pieces of DNA, could replicate in huge quantities independently of chromosomal bacteria DNA and that they could transfer genetic information. It was this process that gave host bacteria the capacity to inherit new genes and therefore new functions such as resistance to antibiotics. Another important tool for creating recombinant DNA was the discovery in the 1960s by the Swiss microbiologist Werner Arber and American biochemist Stuart Linn that bacteria could protect themselves from attack by viruses the production of endonucleases, known as restriction enzymes, which could seek out a single DNA sequence in a virus and cut it precisely in one place. This process prevented the replication of viruses and hence the death of virally infected bacteria. The first restriction enzyme, Escscherichia coli K, was isolated and purified in 1968 by Matthew Meselson and Robert Yuan at Harvard University. Two years later Hamilton O Smith, Thomas Kelly and Kent Welcox at Johns Hopkins University isolated and characterised the first site-specific restriction enzyme, later named HindII. This was demonstrated by Daniel Nathans to be a useful tool for cutting and pasting specific DNA segments. The first protocol for creating recombinant DNA was put forward in the early 1970s by Peter Lobban and Armin Dale Kaiser at Stanford University Medical School. In 1971 Paul Berg, attached to Stanford University, demonstrated the feasibility of splicing and recombining genes for the first time. Two years later, Stanley Cohen and Herbert Boyer, based respectively at Stanford University and University of California at San Francisco, successfully inserted recombinant DNA into bacteria for replication.

Application

Gene cloning has a diverse range of applications. Where it has proven particularly useful has been in mapping out the human genome, the creation of transgenic animals, and the development of insect-resistant crops. It is also pivotal to genetic tests carried out in forensic science and archaeology as well as in tests for determining hereditary disease and paternity. The technology also forms the backbone of hepatitis and human immunodeficiency virus (HIV) diagnostic tests. Recombinant DNA technology has also proven important to the production of vaccines and protein therapies such as human insulin, interferon and human growth hormone. It is also used to produce clotting factors for treating haemophilia and in the development of gene therapy.

Recombinant DNA: timeline of key events

Speigelman was a molecular biologist who investigated how cells form enzymes, DNA and RNA structures. He is credited with improving the nucleic acid hybridisation technique. This technique makes it possible to detect specific DNA and RNA strands in cells. It is now used for analysing the organisation of the genome, studying gene expression and for developing recombinant DNA. 1914-12-14T00:00:00+0000Lederberg was an American geneticist who helped discover the mechanism of genetic recombination in bacteria. This was based on some experiments he performed with Edward Tatum in 1946 which involved mixing two different strains of bacteria. Their experiments also demonstrated for the first time that bacteria reproduced sexually, rather than by cells splitting in two, thereby proving that bacterial genetic systems were similar to those of multicellular organisms. Later on, in 1952, working with Norton Zinder, Lederberg found that certain bacteriophages (viruses that affect bacteria) could carry a bacterial gene from one bacterium to another. In 1958 Lederberg shared the Nobel Prize for Medicine for 'discoveries concerning genetic recombination and the organisation of the genetic material of bacteria.' 1925-05-23T00:00:00+0000Berg was an American biochemist. He first made his name in 1971 by demonstrating it was possible to insert DNA from a bacterium into the a virus' DNA, creating what is called recombinant DNA. This he did as part of his work to study viral chromosomes. He was awarded the Nobel Prize in 1980 for this work. His technique paved the way to the development of genetic engineering and the modern biotechnology industry. Berg was also instrumental in the setting up of the Asilomar Conference on Recombinant DNA, in 1975, which drew up the first guidelines for experiments with genetic engineering. 1926-06-30T00:00:00+0000Ray Wu pioneered the first primer-extension method for DNA sequencing which laid the foundation for the Human Genome Project. He was also instrumental in the application of genetic engineering to agricultural plants to improve their output and resistance to pests, salt and drought. 1928-08-14T00:00:00+0000Nathans was the first scientist to demonstrate how restriction enzymes could be used to cleave DNA and how to piece together its fragments to construct a complete map of DNA. His work inspired the use of restriction enzymes for many different biotechnology applications, including DNA sequencing and the construction of recombinant DNA. He was awarded the Nobel Prize in Physiology or Medicine in 1978 for his work on restriction enzymes. 1928-10-30T00:00:00+0000Werner Arber is a geneticist and microbiologist. He shared the 1978 Nobel Prize in 1978 for helping to discover restriction enzymes and showing their application in molecular genetics. It was based on some work he carried out in the 1960s. Arber indicated in 1965 that restriction enzymes could be used as a tool for cleaving DNA. The enzymes are now an important tool for genetic engineering. 1929-06-03T00:00:00+0000Stahl is a molecular biologist and geneticist who helped to elucidate how DNA is replicated. Together with Matthew Medelsohn, Stahl showed that the double-stranded helix molecule of DNA separates into two strands and that each of these strands serve as a template for the production of a new strand of DNA. They did this in 1958. Following this work, Stahl did extensive work on bacteriophages, viruses that infect bacteria, and their genetic recombination. In 1964 he established that DNA in T4 bacteriophages is circular rather than linear. Eight years later he and his wife, Mary, found a DNA sequence in the lambda bacteriophage necessary to initiate genetic recombination. This laid the foundation for genetic engineering. 1929-10-08T00:00:00+0000Hamilton O Smith is an American microbiologist who helped isolate and characterised the first restriction enzyme from the bacteria Haemophilus influenzae. This he achieved with Kent Wilcox in 1970. They showed that the enzyme degrades foreign phage DNA but not the host's DNA. Now known as HindIII, the restriction enzyme went on to become a major tool for cutting and pasting of specific DNA fragments for the generation of recombinant DNA. Smith was awarded the Nobel Prize for Physiology or Medicine in 1978 for his part in the discovery of the enzyme. In 1995 he and a team at the Institute for Genomic Research completed the DNA sequence of Haemophilus influenzae. It was the first bacterial genome to be deciphered. Later on he helped in the genomic sequencing efforts for the fruit fly and humans at Celera Genomics. 1931-08-23T00:00:00+0000Cohen is an American physician and geneticist whose research has focused on the biology of bacterial plasmids, independent circular units of DNA found in and sometimes exchanged by bacteria. In 1970 he found a way to make Escherichia coli acquire a plasmid that made it resistant to the antibiotic tetracycline. He also discovered with Herbert Boyer a restriction enzyme that could cleave a circular plasmid at a single site. This laid the foundation for their joint experiment in 1973 which demonstrated the feasibility of combining and replicating genetic information from different species. Their experiment involved inserted a gene for frog ribosomal RNA into bacterial cells which then expressed the gene. Three patents were taken out on their technique. These paved the way to the rise of new start-up biotechnology companies, founded on the back of the promise of genetic engineering for generating new therapeutic products. 1935-06-30T00:00:00+0000Together with Stanley Cohen, Boyer demonstrated the possibility of producing recombinant DNA in bacteria in 1973. This they did by combining a gene for frog ribosomal RNA with a bacterial plasmid which was then put into a strain of E-coli for expression. Based on this technique Boyer helped found Genentech, the first biotechnology company dedicated to commercialising recombinant DNA. This he did in 1976 in collaboration with Robert Swanson. 1936-07-10T00:00:00+0000Baltimore shared the 1975 Nobel Prize for his work on the interaction between tumor viruses and the genetic material of the cell. He also spearheaded efforts for the scientific governance of recombinant DNA and genome editing technologies. 1938-03-07T00:00:00+0000Term first used by A. Jost, a Danish microbiologist, in lecture on sexual reproduction in yeast presented to the Technical Institute in Lwow, Poland 1941-01-01T00:00:00+0000Together with Herbert Boyer, Swanson helped found Genentech, the first biotechnology company dedicated to commercialising recombinant DNA. From 1976 to 1990 Swanson was Chief Executive and Director of the company and played an instrumental role in leading it to become the first major biotechnology company to show a profit and go public. 1947-11-29T00:00:00+0000The lambda phage has become a key tool in molecular biology and is important for genetic engineering. It has the advantage that it can be easily grown in E Coli and is not pathogenic except in the case of bacteria. Lederberg's discovery paved the way to understanding the transfer of genetic material between bacteria, the mechanisms involved in gene regulation and how piece of DNA break apart and recombine to make new genes. EM Lederberg, 'Lysogenicity in Escherichia coli strain K-12', Microbial Genetics Bulletin, 1, (1950), 5-9. 1950-01-01T00:00:00+0000Noted by Salvador Luria and his graduate student Mary Human while conducting experiments into the break-up of DNA in phage-infected bateria.1952-01-01T00:00:00+0000The feat was achieved by Arthur Kornberg. He published his experiment in the Journal of Biological Chemsitry in May 1958.1957-10-01T00:00:00+0000Werner Arber, Swiss microbiologist and geneticist, and his doctoral student Daisy Dussoix proposed that bacteria produce restriction and modification enzymes to counter invading viruses. They published their findings in 'Host specificity of DNA produced by Escherichia coli I and II', Journal Molecular Biology, 5 (1962), 18–36 and 37-49.1962-01-23T00:00:00+0000The prediction was published in W. Arber, 'Host-controlled modification of bacteriophage', Annual Review Microbiology, 19 (1965), 365-78. it was based on some research he carried out in the early 1960s with his doctoral student, Daisy Dussoix. They found that bacteria protect themselves against invading viruses by producing two types of enzymes. One cut up the DNA of the virus and the other restricted its growth. Arber believed these two enzymes could provide an important tool for cutting and pasting DNA, the method now used in genetic engineering. 1965-10-01T00:00:00+0000The enzyme was made by four different research teams headed up Martin Gellert, Robert Lehman, Charles Richardson, and Jerard Hurwitz. Its discovery was pivotal to the development of recombinant DNA.1966-01-01T00:00:00+0000Mehran Goulian and Arthur Kornberg managed to assemble the genome using one strand of natural antiviral DNA. The two scientists announced their achievement to a press conference as part of an effort to increase the American public's appreciation of government funded scientific work. It, however, generated debate about whether life should be created in a test tube. The achievement was an important stepping stone to the development of recombinant DNA. 1967-12-14T00:00:00+00001968-01-01T00:00:00+0000This was developed by Peter Lobhan, a graduate student of Dale Kaiser at Stanford University.1969-01-01T00:00:00+0000Achived by Har Gobind Khorana at the University of Wisconsin-Madison1970-01-01T00:00:00+0000The finding was published in Hamilton O Smith, Kent W Wilcox, 'A restriction enzyme from Hemophilus influenzae. I. Purification and general properties',Journal of Molecular Biology, 51/2 (1970), 379-91. Restriction enzymes are now workhorses of molecular biology. They are essential in the development of recombinant DNA and were pivotal to the foundation of the biotechnology industry. 1970-07-01T00:00:00+0000Reverse transcriptase is a restriction enzyme that cuts DNA molecules at specific sites. The enzyme was simultaneously discovered independently by Howard Temin and David Baltimore. Temin made the discovery while working on Rous sacoma virions and Baltimore was working on the poliovirus and vesicular stomatis virus. The discovery laid the foundations for the the disciplines of retrovirology and cancer biology and ability to produce recombinant DNA. The findings were published in D Baltimore, 'RNA-dependent DNA polymerase in virions of RNA tumour viruses' Nature, 226 (1970), 1209–11 and HM Temin, S Mizutani, 'RNA-dependent DNA polymerase in virions of Rous sarcoma virus', Nature, 226 (1970), 1211–13. 1970-07-27T00:00:00+0000The aim of her docrtoal research was to figure out how to replicate and express recombinant DNA in E. coli. 1970-09-01T00:00:00+0000This was done in Dale Kaiser's laboratory by Douglas Berg together with Janet Mertz and David Jackson1971-01-01T00:00:00+0000Robert Pollack contacted Paul Berg to raise concerns about the potential biohazards of experiments Mertz, his doctoral research student, planned to do involving the introduction of genes from the oncovirus SV40 in the human gut bacteria, E. Coli. Following this Berg self-imposed a moratorium on experiments in his laboratory involving the cloning of SV40 in E-Coli.1971-06-01T00:00:00+0000The power of restriction enzymes to cut DNA was demonstrated by Kathleen Danna, a graduate student, with Daniel Nathans, her doctoral supervisor, at Johns Hopkins University. They published the technique in 'Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae', PNAS USA, 68/12 (1971), 2913-17.1971-12-01T00:00:00+0000This took place during an unscheduled extra session held one evening during a three-day EMBO workshop near Basel on DNA restriction and modification. The session was chaired by Norton Zinder. The discussion set the stage for the subsequent Asilomar Conference in 1975 which led to the first guideline for experiments with genetic engineering. 1972-09-26T00:00:00+0000The recombinant DNA was made by Paul Berg and colleagues. It was generated by cutting DNA with a restriction and then using ligase to paste together two DNA strands to form a hybrid circular molecule. The method was published in D A Jackson, R H Symons, P Berg, 'Biochemical Method for Inserting New Genetic Information into DNA of Simian Virus 40: Circular SV40 DNA Molecules Containing Lambda Phage Genes and the Galactose Operon of Escherichia coli', PNAS USA, 69/10 (1972), 2904-09.1972-10-01T00:00:00+0000It was based on their finding that when DNA is cleaved with EcoRI, a restriction enzyme, it has sticky ends. JE Mertz, RW Davis, 'Cleavage of DNA by RI restriction endonuclease generates cohesive ends', PNAS, 69, 3370–3374 (1972). 1972-11-01T00:00:00+0000The work was carried out by Stanley Cohen and Annie Chang at Stanford University in collaboration with Herbert Boyer and Robert Helling at the University of California San Francisco. They managed to splice sections of viral DNA and bacterial DNA with the same restriction enzyme to create a plasmid with dual antibiotic resistance. They then managed to insert this recombinant DNA molecule into the DNA of bacteria to express the new recombinant DNA. The technique showed it was possible to reproduce recombinant DNA in bacteria. It was published in SN Cohen, ACY Chang, HW Boyer, RB Belling, 'Construction of Biologically Functional Bacterial Plasmids In Vitro', PNAS USA, 10/11 (1973), 3240-3244. 1973-11-01T00:00:00+0000The National Institutes of Health forms a Recombinant DNA Advisory Committee to oversee recombinant genetic research.1974-01-01T00:00:00+0000JF Morrow, SN Cohen, ACY Chang, HW Boyer, HM Goodman, RB Helling, 'Replication and Transcription of Eukaryotic DNA in Esherichia coli', PNAS USA, 171/5 (1974), 1743-47.1974-05-01T00:00:00+0000The call was published by P Berg et al 'Biohazards of Recombinant DNA,' Science, 185 (1974), 3034. It argued for the establishment of an advisory committee to oversee experimental procedures to evaluate the potential biological hazards of recombinant DNA molecules and develop procedures to minimise the spread of such molecules within human and other populations. 1974-07-05T00:00:00+0000Her thesis focused on methods to isolate and characterise mutant variants of SV40 1975-01-01T00:00:00+0000The conference, organised by Paul Berg had 140 professional participants (including biologists, physicians and lawyers). In addition to the moratorium the conference established several principles for safely conducting any genetic engineering. Containment was considered essential to any experimental design, such as the use of hoods, and the use of biological barriers was suggested to limit the spread of recombinant DNA. This included using bacterial hosts that could not survive in natural environment and the use of vectors (plasmids, bacteriophages and other viruses) that could only grow in specified hosts. The conference also called for a moratorium on genetic engineering research in order to have time to estimate the biohazard risks of recombinant DNA research and develop guidelines.1975-02-01T00:00:00+0000Yeast genes expressed in E. coli bacteria for the first time1976-01-01T00:00:00+0000Robert Swanson, venture capitalist and Herbert Boyer, American biochemist, established Genentech in San Francisco. It was the first biotechnology company established specifically dedicated to commercialising recombinant DNA. Its founding marked the start of what was to become a burgeoning biotechnology industry. 1976-04-01T00:00:00+0000The guidelines were issued following a public meeting held in February 1976. 1976-06-23T00:00:00+0000Genetically engineered bacteria are used to synthesize human growth protein.1977-01-01T00:00:00+0000Genentech scientists succeed in genetically engineering human insulin in E-Coli.1978-01-01T00:00:00+0000The prize was jointly awarded to Werner Arber, Daniel Nathans and Hamilton O Smith. Arber was the first to discover the enzymes; Smitth demonstrated their capacity to cut DNA at specific sites and Nathans showed how they could be used to construct genetic maps. With their ability to cut DNA into defined fragments restriction enzymes paved the way to the development of genetic engineering. 1978-10-01T00:00:00+0000The patent was filed on the basis of work undertaken by Kenneth Murray. 1978-12-22T00:00:00+0000The cloning, achieved by Beverly Griffin with Tomas Lindahl, was announced to a meeting at Cold Spring Harbor1979-01-01T00:00:00+0000The work, funded by Biogen, was undertaken as part of a project to develop recombinant hepatitis B vaccine. It was published in CJ Burrell, P Mackay, PJ Greenaway, PH Hofsneider, K Murray, 'Expression in Escheria Coli of hepatitis B virus DNA sequences cloned in plasmid pBR322', Nature, 279/5708 (1979), 43-47. 1979-02-01T00:00:00+0000F Galibert, E Mandart, F Fitoussi, P Tiollais, P Charnay, , 'Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature, 281/5733 (1979), 646-50; P. Charnay, C Pourcel, A Louise, A Fritsch, P Tiollais, 'Cloning in Escherichia coli and physical structure of hepatitis B virion DNA', PNAS USA, 76/5 (1979), 2222-26; P Charnay, E Mandart, A Hampe, F Fitoussi, P Tiollais, F Galibert, 'Localization on the viral genome and nucleotide sequence of the gene coding for the two major polypeptides of the hepatitis B surface antigen (HBs Ag)', Nucleic Acids Research, 7/2 (1979), 335-46.1979-05-01T00:00:00+0000The research was funded by Merck with the aim of developing a recombinant vaccine against hepatitis B. It was published in P Valenzuela, P Gray, M Quiroga, J Zaldivar, H M Goodman, WJ Rutter, 'Nucleotide sequence of the gene coding for the major protein of hepatitis B virus surface antigen', Nature, 280/5725 (1979), 815e819.1979-08-30T00:00:00+0000The patent was based on the work of Kenneth Murray. It was granted in July 1990 as European Patent (UK) No 0182442. 1979-12-21T00:00:00+0000US Supreme Court, in the landmark case Diamond v. Chakrabarty, approves the principle of patenting genetically engineered life forms1980-01-01T00:00:00+0000The American scientists Stanley Cohen and Herbert Boyer are awarded the first US patent for gene cloning.1980-01-01T00:00:00+0000Milstein suggests at a Wellcome Foundation lecture that by using genetic engineering scientists might be able to design tailor-made monoclonal antibodies that mimic antibodies made by the human body. This would free them up from a dependence on rodents for producing monoclonal antibodies. He publishes the idea in C. Milstein, 'Monoclonal antibodies from hybrid myelomas: Wellcome Foundation Lecture 1980', Proceedings Royal Society of London, 211 (1981), 393-412.1980-01-01T00:00:00+0000JC Edman, P Gray, P Valenzuela, LB Rall, WJ Rutter, 'Integration of hepatitis B virus sequences and their expression in a human hepatoma cell', Nature, 286/5772 (1980), 535-38.1980-07-31T00:00:00+0000The mice were made with the help recombinant DNA technology. JW Gordon, GA Scangos, DJ Plotkin, J A Barbosa, FH Ruddle, 'Genetic transformation of mouse embryos by microinjection of purified DNA', PNAS USA, 77 (1980), 7380–4.1980-09-01T00:00:00+0000First genetically-engineered plant is reported1981-01-01T00:00:00+0000First mice genetically cloned1981-01-01T00:00:00+00001981-07-01T00:00:00+0000The work, led by Beverly Griffin, opened up the possibility of sequencing the virus. It was published in J R Arrand, L. Rymo, J E Walsh, E Bjorck, T Lindahl and B E Griffin, ‘Molecular cloning of the complete Epstein-Barr virus genome as a set of overlapping restriction endonuclease fragments’, Nucleic Acids Research, 9/13 (1981), 2999-2014.1981-07-10T00:00:00+0000The first drug (human insulin), based on recombinant DNA, is marketed. 1982-10-01T00:00:00+0000Speigelman was an American molecular biologist who investigated how cells form enzymes, DNA and RNA structures. He is credited with improving the nucleic acid hybridisation technique. This technique makes it possible to detect specific DNA and RNA strands in cells. It is now used for analysing the organisation of the genome, studying gene expression and for developing recombinant DNA.1983-01-20T00:00:00+0000The trial was done with 37 healthy adult volunteers. The vaccine was made using HBsAg cloned in yeast. EM Scolnick, AA McLean, DJ West, WJ McAleer WJ Miller, EB Buynak, 'Clinical evaluation in healthy adults of a hepatitis B vaccine made by recombinant DNA', JAMA 251/21 (1984), 2812-15. 1984-06-01T00:00:00+0000Two teams of scientists publish methods for the generation of chimeric monoclonal antibodies, that is antibodies possessing genes that are half-human and half mouse. Each team had developed their techniques separate from each other. The first team was lead by Michael Neuberger together with Terence Rabbitts and other colleagues at the Laboratory of Molecular Biology, Cambridge. The second team consisted of Sherie Morrison and colleagues at Stanford University together with Gabrielle Boulianne and others at the University of Toronto. 1984-12-01T00:00:00+0000Greg Winter together with other colleagues from the Laboratory Molecular Biology demonstrate the feasibility of building a new more human-like monoclonal antibody by grafting on to the humab antibody portions of a variable region from a mouse antibody. This reduced the mouse component of the monoclonal antibody to just 5%, making the monoclonal antibody safer and more effective for use in humans. The technique was published in PT Jones, PH Dear, J Foote, MS Neuberger, G Winter, 'Replacing the complementarity-determining regions in a mouse antibody with those from a mouse', Nature, 321 (29 May 1986), 522-5.1986-05-01T00:00:00+0000The vaccine was first approved in West Germany, in May, and then in the US in July. The vaccine was regarded as a breakthrough because it was made from a genetically engineered sub-particle of the virus. This made it much safer than the original vaccine which used the virus sub-particle sourced from the blood of hepatitis B sufferers. The vaccine heralded a new era for the production of vaccines and is a major weapon against one of the most infectious diseases. 1986-05-01T00:00:00+0000Hoffmann-LaRoche and Schering-Plough gain FDA permission to market genetically engineered alpha interferon for use as treatment hairy cell leukaemia. The development of interferon rested on the application of both genetic cloning and monoclonal antibodies. 1986-06-04T00:00:00+00001986-12-01T00:00:00+0000JH Hoofnagle, KD Mullen, B Jones, et al, 'Treatment of chornic non-A, non and non-B hepatitis with recombinant human alpha interferon' NEJM, 315 (1986), 1575-78.1986-12-18T00:00:00+0000Campath-1G is humanised, resulting in Campath-1H. It is accomplished with technology developed by Greg Winter.1988-01-01T00:00:00+0000USPTO patent 4,736,866 awarded for transgenic mouse with activated oncogenes created by Philip Leder and Timonthy A Stewart at Harvard University. The two scientists isolated a gene that causes cancer in many mammals, including humans, and inserted it into fertilised mouse eggs. The aim was to genetically engineer a mouse as a model for furthering cancer research and the testing of new drugs. It was the first animal ever given patent protection in the USA. 1988-04-12T00:00:00+00001989-01-01T00:00:00+00001989-05-01T00:00:00+0000The drug was developed by Schering Plough. The drug helps suppress the replication of the hepatitis B virus. 1992-07-13T00:00:00+0000Cetus was the first biotechnology company created. It was set up in California by Ronald E. Cape, Peter Farley, and Nobelist Donald A. Glaser. Cetus Corporation initially focused its efforts on the automation of selecting for industrial microorganisms that could produce greater amounts of chemical feedstocks, antibiotics or vaccine components. From the late 1970s the company turned its attention to genetic engineering and by 1983 had created its own recombinant interleukin (IL-2) for treating renal cancer, which was eventually approved 2 years after Cetus was sold. The company is best known for its development of development of the revolutionary DNA amplification technique known as polymerase chain reaction (PCR) technology. 1993-10-13T00:00:00+0000The drug, a recombinant human deoxyribonuclease, was developed by the Genentech researcher Steven Shak. It was the first new treatment for cystic fibrosis in 30 years. The enzyme was engineered to dissolve mucus plugs in the lungs of cystic fibrosis patients. The product was marketed as Pulmozyme. 1993-12-30T00:00:00+0000Abciximab (ReoPro) approved by the FDA and European regulatory authorities to prevent blot clots during coronary artery procedures like angioplasty. The monoclonal antibody was originally developed by Barry Coller at State University of New York and commercially developed by Centocor. The drug showed for the first time that monoclonal antibodies could be used for the treatment of acute disease conditions. 1994-12-22T00:00:00+0000Daclizumab was approved by the FDA for the prevention of acute rejection of kidney transplants. The monoclonal antibody was developed by Protein Design Labs using a humanising method devised by Cary Queen and marketed together with F. Hoffmann-La Roche. 1997-12-01T00:00:00+0000Nathans was the first scientist to demonstrate how restriction enzymes could be used to cleave DNA and how to piece together its fragments to construct a complete map of DNA. His work inspired the use of restriction enzymes for many different biotechnology applications, including DNA sequencing and the construction of recombinant DNA. He was awarded the Nobel Prize in Physiology or Medicine in 1978 for his work on restriction enzymes.1999-11-16T00:00:00+0000Together with Herbert Boyer, Swanson helped found Genentech, the first biotechnology company dedicated to commercialising recombinant DNA. From 1976 to 1990 Swanson was Chief Executive and Director of the company and played an instrumental role in leading it to become the first major biotechnology company to show a profit and go public. 1999-12-06T00:00:00+0000Lederberg was an American geneticist who helped discover the mechanism of genetic recombination in bacteria. This was based on some experiments he performed with Edward Tatum in 1946 which involved mixing two different strains of bacteria. Their experiments also demonstrated for the first time that bacteria reproduced sexually, rather than by cells splitting in two, thereby proving that bacterial genetic systems were similar to those of multicellular organisms. Later on, in 1952, working with Norton Zinder, Lederberg found that certain bacteriophages (viruses that affect bacteria) could carry a bacterial gene from one bacterium to another. In 1958 Lederberg shared the Nobel Prize for Medicine for 'discoveries concerning genetic recombination and the organisation of the genetic material of bacteria.' 2008-02-02T00:00:00+0000Ray Wu pioneered the first primer-extension method for DNA sequencing which laid the foundation for the Human Genome Project. He was also instrumental in the application of genetic engineering to agricultural plants to improve their output and resistance to pests, salt and drought. 2008-02-10T00:00:00+0000Twelve patients with HIV treated between 2009 and 2014 report benefits from genetically engineered virus with a rare mutatiuon known to protect against HIV (CCR5 deficiency).2014-03-01T00:00:00+0000Berg was an American biochemist. He first made his name in 1971 by demonstrating it was possible to insert DNA from a bacterium into the a virus' DNA, creating what is called recombinant DNA. This he did as part of his work to study viral chromosomes. He was awarded the Nobel Prize in 1980 for this work. His technique paved the way to the development of genetic engineering and the modern biotechnology industry. Berg was also instrumental in the setting up of the Asilomar Conference on Recombinant DNA, in 1975, which drew up the first guidelines for experiments with genetic engineering. 2023-02-15T00:00:00+0000
Date Event People Places
14 Dec 1914Solomon Spiegelman was born in Brooklyn, NY, USASpiegelmanUniversity of Minnesota
23 May 1925Joshua Lederberg was born in Montclair, NJ, USAJoshua LederbergUniversity of Wisconsin
30 Jun 1926Paul Berg was born in New York NY, USABergStanford University
14 Aug 1928Ray Wu was born in Beijing, ChinaWuCornell University
30 Oct 1928Daniel Nathans was born in Wilmington, Delaware, USANathansJohns Hopkins University
3 Jun 1929Werner Arber was born in Granichen, SwitzerlandArberUniversity of Geneva
8 Oct 1929Franklin W Stahl was born in Boston, Massachusetts, USAStahl California Institute of Technology, University of Missouri, University of Oregon
23 Aug 1931Hamilton O Smith was born in New York City, USASmithJohns Hopkins University, Celera
30 Jun 1935Stanley Norman Cohen was born in Perth Amboy, NJ, USACohenStanford University
10 Jul 1936Herbert Boyer was born in Derry, Pennsylvania, USABoyer University of California San Francisco, Genentech
7 Mar 1938David Baltimore was born in New York CityBaltimoreNew York City
1941Term 'genetic engineering' first coinedJost 
29 Nov 1947Robert Swanson was born in Florida, USASwansonGenentech
January 1950Esther Lederberg discovered the lambda phageEsther LederbergUniversity of Wisconsin
1952First observation of the modification of viruses by bacteriaLuria, HumanUniversity of Illinois
October 1957First synthesis of DNA in a test tubeKornbergWashington University in St. Louis
23 Jan 1962Idea of restriction and modification enzymes bornArber, DussoixUniversity of Geneva
1 Oct 1965Werner Arber predicted restriction enzymes could be used as a labortory tool to cleave DNAArberUniversity of Geneva
1966Discovery ligase, an enzyme that facilitates the joining of DNA strandsGellert, Lehman, Richardson, Hurwitz 
14 Dec 1967Functional, 5,000-nucleotide-long bacteriophage genome assembledGoulian, KornbergStanford University, Chicao University
1968Paul Berg started experiments to generate recombinant DNA moleculesBergStanford University
1969New idea for generating recombinant DNA conceivedLobhanStanford University
1970First complete gene synthesised KhoranaUniversity of Wisconsin
July 1970First restriction enzyme isolated and characterisedSmith, WilcoxJohns Hopkins University
27 Jul 1970Reverse transcriptase first isolatedBaltimore, Temin, MizutaniMassachusetts Institute of Technology, University of Wisconsin
September 1970Mertz started her doctorate in biochemistry at Stanford University under Paul BergMertz, Berg 
1971First plasmid bacterial cloning vector constructedBerg, Mertz, JacksonStanford University
June 1971Janet Mertz forced to halt experiment to clone recombinant DNA in bacteria after safety concerns raisedMertz, Berg, PollackStanford University
December 1971First experiments published demonstrating the use of restriction enzymes to cut DNADanna, NathansJohns Hopkins University
26 Sep 1972 - 4 Sep 1972First time possible biohazards of recombinant DNA technology publicly discussedZinderEMBO
1 Oct 1972First recombinant DNA generatedBerg, Jackson, SymonsStanford University
November 1972Janet Mertz and Ronald Davis published first easy-to-use technique for constructing recombinant DNA showed that when DNA is cleaved with EcoRI, a restriction enzyme, it has sticky endsMertz, DavisStanford University
1 Nov 1973First time DNA was successfully transferred from one life form to anotherCohen, Chang, BoyerStanford University, University of California San Francisco
1974Regulation begins for recombinant genetic research 
1 May 1974Recombinant DNA successfuly reproduced in Escherichia coliMarrow, Cohen, Chang, Boyer, Goodman, HellingStanford University, University of California San Francisco
July 1974Temporary moratorium called for on genetic engineering until measures taken to deal with potential biohazardsBerg, Baltimore, Boyer, Cohen 
January 1975Mertz completed her doctorate MertzStanford University
February 1975Asilomar Conference called for voluntary moratorium on genetic engineering researchBerg 
1976Yeast genes expressed in E. coli bacteria for the first time 
April 1976Genentech foundedSwanson, BoyerGenentech Inc
23 Jun 1976NIH released first guidelines for recombinant DNA experimentation 
1977Human growth hormone genetically engineered 
1978Human insulin produced in E-coliGenentech
October 1978Nobel Prize given in recognition of discovery of restriction enzymes and their application to the problems of molecular geneticsArber, Nathans, SmithJohns Hopkins University, University of Geneva
December 1978Biogen filed preliminary UK patent for technique to clone hepatitis B DNA and antigensKenneth MurrayBiogen, University of Edinburgh
1979First DNA fragments of Epstein Barr Virus cloned Griffin, LindahlImperial Cancer Research Fund Laboratories, University of Gothenberg
February 1979University of Edinburgh scientists published the successful isolation and cloning DNA fragments of the hepatitis B virus in Escherichia coliBurrell, Mackay, Greenaway, Hofschneider, K MurrayUniversity of Edinburgh, Microbiological Research Establishment, Biogen
May 1979 - Oct 1979Pasteur Institute scientists reported successful cloning of hepatitis B DNA in Escherichia coliGalibert, Mandart, Fitoussi, Tiollais, Charnay, HampePasteur Institute
30 Aug 1979UCSF scientists announced the successful cloning and expression of HBsAg in Escherichia coliValenzuela, Gray, Quiroga, Zaldivar, Goodman, RutterUniversity of California San Francisco, Merck
21 Dec 1979Biogen applied for European patent to clone fragment of DNA displaying hepatitis B antigen specificityMurrayBiogen
1980Genetic engineering recognised for patenting 
1980First patent awarded for gene cloningCohen, BoyerStanford University Medical School
1980Cesar Milstein proposed the use of recombinant DNA to improve monoclonal antibodiesMilsteinLaboratory of Molecular Biology
31 Jul 1980UCSF scientists published method to culture HBsAg antigens in cancer cellsEdman, Gray, Valenzuela, Rall, RutterUniversity of California San Francisco
September 1980Scientists reported the first successful development of transgenic miceBarbosa, Gordon, Plotkin, Ruddle, ScangosYale University
1981First genetically-engineered plant reported 
1981First genetically cloned mice 
July 1981UCSF and Merck filed patent to snthesise HBsAg in recombinant yeastRutterUniversity of California San Francisco, Merck
10 Jul 1981Complete library of overlapping DNA fragments of Epstein Barr Virus clonedGriffin, Arrand, Walsh, Bjorck, RymoImperial Cancer Research Fund Laboratories, University of Gothenberg
October 1982First recombinant DNA based drug approvedGenentech Inc
20 Jan 1983Solomon Spiegelman diedSpiegelmanUniversity of Minnesota
1 Jun 1984Genetically engineered vaccine against hepatitis B reported to have positive trial resultsScolnick, McLean, West, McAleer , Miller, BuynakMerck, University California San Francisco
1984First chimeric monoclonal antibodies developed, laying foundation for safer and more effective monoclonal antibody therapeuticsNeuberger, Rabbitts, Morrison, Oi, Herzenberg, Boulianne, Schulman, HozumiLaboratory of Molecular Biology, Stanford Univerity Medical School
May 1986First humanised monoclonal antibody createdDear, Foote, Jones, Neuberger, WinterLaboratory of Molecular Biology
1986First genetically engineered vaccine against hepatitis B approvedScolnickMerck
June 1986Interferon approved for treating hairy cell leukaemia 
December 1986Genetically engineered hepatitis B vaccine, Engerix-B, approved in BelgiumSmithKline Biologicals
18 Dec 1986Results released from first small-scale clinical trial of recombinant interferon-alpha therapy for post-transfusion chronic hepatitis BHoofnagle, Mullen, Jones, Rustgi, Di Bisceglie, Peters, Waggoner, ParkNational Institutes of Health
1988Campath-1H is created - the first clinically useful humanised monoclonal antibody.Winter, Waldmann, Reichmann, ClarkCambridge University, Laboratory of Molecular Biology
12 Apr 1988OncoMouse patent grantedLeder, StewartHarvard University
January 1989Genetically engineered hepatitis B vaccine, Engerix-B, approved in USSmithKline Biologicals
May 1989Genetically engineered hepatitis B vaccine, GenHevac, approved in FrancePasteur Vaccins
13 Jul 1992FDA approved the use of genetically engineered interferon-alpha, Intron A, for the treatment of hepatitis BSchering-Plough
13 Oct 1993Cetus Corporation was sold to Chiron and its patent rights sold for US$300 million to Hoffman-La RocheCape, Farley, Glaser MullisCetus Corporation, Chiron, Hoffman-La Roche
30 Dec 1993FDA appproved genetically engineered enzyme drug for cystic fibrosisSnakGenentech
22 Dec 1994First chimeric monoclonal antibody therapeutic approved for marketColler, SchoemakerCentocor, State University of New York
December 1997First humanised monoclonal antibody approved for marketQueenProtein Design Labs, Roche
16 Nov 1999Daniel Nathans diedNathans Johns Hopkins University
6 Dec 1999Robert Swanson diedSwansonGenentech
2 Feb 2008Joshua Lederberg diedJoshua LederbergUniversity of Wisconsin
10 Feb 2008Ray Wu died in Ithaca, USAWuCornell University
March 2014Promising results announced from trial conducted with HIV patients 
15 Feb 2023Paul Berg diedBergStanford University

14 Dec 1914

Solomon Spiegelman was born in Brooklyn, NY, USA

23 May 1925

Joshua Lederberg was born in Montclair, NJ, USA

30 Jun 1926

Paul Berg was born in New York NY, USA

14 Aug 1928

Ray Wu was born in Beijing, China

30 Oct 1928

Daniel Nathans was born in Wilmington, Delaware, USA

3 Jun 1929

Werner Arber was born in Granichen, Switzerland

8 Oct 1929

Franklin W Stahl was born in Boston, Massachusetts, USA

23 Aug 1931

Hamilton O Smith was born in New York City, USA

30 Jun 1935

Stanley Norman Cohen was born in Perth Amboy, NJ, USA

10 Jul 1936

Herbert Boyer was born in Derry, Pennsylvania, USA

7 Mar 1938

David Baltimore was born in New York City

1941

Term 'genetic engineering' first coined

29 Nov 1947

Robert Swanson was born in Florida, USA

Jan 1950

Esther Lederberg discovered the lambda phage

1952

First observation of the modification of viruses by bacteria

Oct 1957

First synthesis of DNA in a test tube

23 Jan 1962

Idea of restriction and modification enzymes born

1 Oct 1965

Werner Arber predicted restriction enzymes could be used as a labortory tool to cleave DNA

1966

Discovery ligase, an enzyme that facilitates the joining of DNA strands

14 Dec 1967

Functional, 5,000-nucleotide-long bacteriophage genome assembled

1968

Paul Berg started experiments to generate recombinant DNA molecules

1969

New idea for generating recombinant DNA conceived

1970

First complete gene synthesised

Jul 1970

First restriction enzyme isolated and characterised

27 Jul 1970

Reverse transcriptase first isolated

Sep 1970

Mertz started her doctorate in biochemistry at Stanford University under Paul Berg

1971

First plasmid bacterial cloning vector constructed

Jun 1971

Janet Mertz forced to halt experiment to clone recombinant DNA in bacteria after safety concerns raised

Dec 1971

First experiments published demonstrating the use of restriction enzymes to cut DNA

26 Sep 1972 - 4 Sep 1972

First time possible biohazards of recombinant DNA technology publicly discussed

1 Oct 1972

First recombinant DNA generated

Nov 1972

Janet Mertz and Ronald Davis published first easy-to-use technique for constructing recombinant DNA showed that when DNA is cleaved with EcoRI, a restriction enzyme, it has sticky ends

1 Nov 1973

First time DNA was successfully transferred from one life form to another

1974

Regulation begins for recombinant genetic research

1 May 1974

Recombinant DNA successfuly reproduced in Escherichia coli

Jul 1974

Temporary moratorium called for on genetic engineering until measures taken to deal with potential biohazards

Jan 1975

Mertz completed her doctorate

Feb 1975

Asilomar Conference called for voluntary moratorium on genetic engineering research

1976

Yeast genes expressed in E. coli bacteria for the first time

Apr 1976

Genentech founded

23 Jun 1976

NIH released first guidelines for recombinant DNA experimentation

1977

Human growth hormone genetically engineered

1978

Human insulin produced in E-coli

Oct 1978

Nobel Prize given in recognition of discovery of restriction enzymes and their application to the problems of molecular genetics

Dec 1978

Biogen filed preliminary UK patent for technique to clone hepatitis B DNA and antigens

1979

First DNA fragments of Epstein Barr Virus cloned

Feb 1979

University of Edinburgh scientists published the successful isolation and cloning DNA fragments of the hepatitis B virus in Escherichia coli

May 1979 - Oct 1979

Pasteur Institute scientists reported successful cloning of hepatitis B DNA in Escherichia coli

30 Aug 1979

UCSF scientists announced the successful cloning and expression of HBsAg in Escherichia coli

21 Dec 1979

Biogen applied for European patent to clone fragment of DNA displaying hepatitis B antigen specificity

1980

Genetic engineering recognised for patenting

1980

First patent awarded for gene cloning

1980

Cesar Milstein proposed the use of recombinant DNA to improve monoclonal antibodies

31 Jul 1980

UCSF scientists published method to culture HBsAg antigens in cancer cells

Sep 1980

Scientists reported the first successful development of transgenic mice

1981

First genetically-engineered plant reported

1981

First genetically cloned mice

Jul 1981

UCSF and Merck filed patent to snthesise HBsAg in recombinant yeast

10 Jul 1981

Complete library of overlapping DNA fragments of Epstein Barr Virus cloned

Oct 1982

First recombinant DNA based drug approved

20 Jan 1983

Solomon Spiegelman died

1 Jun 1984

Genetically engineered vaccine against hepatitis B reported to have positive trial results

1984

First chimeric monoclonal antibodies developed, laying foundation for safer and more effective monoclonal antibody therapeutics

May 1986

First humanised monoclonal antibody created

1986

First genetically engineered vaccine against hepatitis B approved

Jun 1986

Interferon approved for treating hairy cell leukaemia

Dec 1986

Genetically engineered hepatitis B vaccine, Engerix-B, approved in Belgium

18 Dec 1986

Results released from first small-scale clinical trial of recombinant interferon-alpha therapy for post-transfusion chronic hepatitis B

1988

Campath-1H is created - the first clinically useful humanised monoclonal antibody.

12 Apr 1988

OncoMouse patent granted

Jan 1989

Genetically engineered hepatitis B vaccine, Engerix-B, approved in US

May 1989

Genetically engineered hepatitis B vaccine, GenHevac, approved in France

13 Jul 1992

FDA approved the use of genetically engineered interferon-alpha, Intron A, for the treatment of hepatitis B

13 Oct 1993

Cetus Corporation was sold to Chiron and its patent rights sold for US$300 million to Hoffman-La Roche

30 Dec 1993

FDA appproved genetically engineered enzyme drug for cystic fibrosis

22 Dec 1994

First chimeric monoclonal antibody therapeutic approved for market

Dec 1997

First humanised monoclonal antibody approved for market

16 Nov 1999

Daniel Nathans died

6 Dec 1999

Robert Swanson died

2 Feb 2008

Joshua Lederberg died

10 Feb 2008

Ray Wu died in Ithaca, USA

Mar 2014

Promising results announced from trial conducted with HIV patients

15 Feb 2023

Paul Berg died

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