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CRISPR Timeline | Broad Institute

CRISPR Timeline

CRISPR Timeline | Berkeley News


Timeline of CRISPR (

A Timeline of the CRISPR/Cas System

A Timeline of the CRISPR/Cas System | Today's Clinical Lab

CRISPR-Cas9: Timeline of key events

Timeline: CRISPR-Cas9 (

CRISPR History and Development for Genome Engineering

Addgene: CRISPR History and Development for Genome Engineering

Human Gene Editing: A Timeline of CRISPR Cover Stories

Human Gene Editing: A Timeline of CRISPR Cover Stories | Center for Genetics and Society

A Brief History of CRISPR-Cas9 Genome-Editing Tools

A Brief History of CRISPR-Cas9 Genome-Editing Tools (

The CRISPR Patent Battle Is Finally Over

The CRISPR Patent Battle Is Finally Over | Brownstone Research


1859 Charles Darwin publishes The Origin of the Species
1866 Gregor Mendel discovers the basic principles of genetics - he postulated a set of rules to explain the inheritance of biological characteristics in living organisms
1869 Frederich Miescher identifies nuclein
1900s the eugenics movement
1900 Mendel’s theories are rediscovered by researchers
1902 Sir Archibald Garod is the first to associate Mendel’s theories with a human disease
1903 Sutton postulates that genes are located on chromosomes
1910 Morgan’s experiments prove genes are located
on chromosomes
1941 term genetic engineering first coined
1944 Oswald Avery identifies DNA as the transforming principle - Avery, Mcleod, and McCarty demonstrated that genes are composed of DNA rather than protein
1950 discovered plasmids, small module pieces of DNA could replicate in huge quantities independently of chromosomes in bacterial DNA and that they could transfer genetic information
1950 Erwin Chargaff discovers that DNA composition
is species specific
January 1950 Esther Lederberg discovered the lambda phage
1952 Rosalind Franken photographs crystallized DNA fibers
1952 Hershey and Chase confirm role of DNA as the basic
genetic material
1952 first observation of the modification of viruses by bacteria
1953 James Watson and Francis Crick discover the double helix structure of DNA - structure of DNA is determined
1953 George Gamow and the RNA tie club
October 1957 first synthesis of DNA in a test tube
1959 an additional copy of chromosome linked to Downs syndrome

1960 genetic code deciphered
1960 Swiss microbiologists Werner Arber and American biochemist Stuart Linn discover that bacteria can protect themselves from attack by viruses by producing endonucleases known as restriction enzymes, which can seek out a single DNA sequence in a virus and cut it precisely in one place = prevents the replication of viruses and the death of virally infected bacteria
Late 1960s recombinant DNA first breakthrough of rDNA = with discovery of restriction and endonucleases and restriction enzyme by Werner Arber and Hamilton Smith
the restriction enzymes were discovered in microorganism - the enzymes protect the host cell from bacteriophage
January 23, 1962 idea of restriction and modification enzymes born
October 1, 1965 Werner Aber predicted restriction enzymes could be used as a laboratory tool to cleave DNA
1966 discovery of ligase, an enzyme that facilitates the joining of DNA strands
December 14, 1967 functional, 5000-nucleotide-long bacteriophage genome assembled
1968 the first restriction enzyme Escherichia coli K was isolated and purified by Matthew Meselson and Robert Yuan at Harvard
1968 Paul Berg started experiments to generate recombinant
DNA molecules
1969 new idea for generating recombinant DNA conceived
1969 Herbert Boyer isolated restriction enzyme EcoRI from E. coli that cleaves the DNA between G and A in the base sequence GAATTC
1969 Paul Berg and Peter Lobban independently conceive an approach to create rDNAs in vitro and use them to manipulate genes across species
early 1970s first protocol for creating recombinant DNA was put forward by Peter Lobba and Armin Dale Kaiser at Stanford University medical school
1970 Howard Temin and Davin Baltimore discovered the enzyme reverse transcriptase from retrovirauses; later on this enzyme was used to construct a DNA called complementary DNA (cDNA) from any mRNA
1970 first complete gene synthesized
1970 Hamilton O Smith and Thomas Kelly and Kent Welcox at Johns Hopkins University isolated and characterized the first site-specific restriction enzymes later named HANDLL

July 1970 first restriction enzyme isolated and characterized
July 27, 1970 reverse transcriptase first isolated
1971 Paul Berg at Stanford University demonstrated the feasibility of splicing and recombining genes for the first time - Cohen and Boyer develop techniques for rDNA technology, to allow transfer of genetic material from one organism to another
1971 first plasmid bacterial cloning vector constructed
June 1971 Janet Mertz forced to halt experiment to clone recombinant DNA in bacteria after safety concerns raised
December 1971 first experiments published demonstrating the use of restriction enzymes to cut DNA
1971 Douglas Berg and colleagues isolate the first plasmid bacterial cloning vector
1971 Robert Pollack raises first concerns about potential biohazards of cloning
1971 David Jackson and colleagues, Peter Lobban, and A.D. Kaiser develop the method for joining DNAs in vitro
1972 David Jackson, Robert Symons and Paul Berg successfully generated rDNA molecules; they allowed the sticky ends of complementary DNA by using an enzyme DNA ligase
1972 Jackson and colleagues create the first chimeric DNA in vitro
1972 Janet Mertz and Ronald Davis discover a new approach to create specific chimeric DNAs in vitro
1972 Stanley Cohen and colleagues isolate a new cloning vector, pSC101, and create bacterial intra-and interspecies rDNAs
September 1972 first time possible biohazards of recombinant DNA technology publicly discussed
October 1, 1972 first recombinant DNA generated
November 1972 Janet Mertz and Ronald Davis published first EZ-2-use technique for constructing recombinant DNA; showed that when DNA is cleaved with EcoRI, a restriction enzyme, it has sticky ends
1973 for first time S. Cohen and H. Boyer developed a recombinant plasmid which after using as a vector replicated well within a bacterial host
1973 John Morrow and colleagues clone and propagate ribosomal DNA genes from a eukaryote in E. coli
1973 first gene, for insulin production, cloned,
using rDNA technology
March 6, 1973 Genentech startup
November 1, 1973 first time DNA was successfully transferred from one life form to another
1974 regulation begins for recombinant genetic research
1974 first expression in bacteria of a gene cloned from
a different species
May 1, 1974 recombinant DNA successfully reproduced in Escherichia coli
July 1974 temporary moratorium called for on genetic engineering until measures taken to deal with potential biohazards
1975 Edwin M. Southern developed a method for detection of specific DNA fragments for isolation of a gene from complex mixture of DNA; this method is known as the Southern blotting technique
February 1975 Asilomar conference called for voluntary moratorium on genetic engineering research
1976 first prenatal diagnosis by using gene specific probe
1976 yeast genes expressed in E. coli bacteria for the first time
April 1976 first new biotechnology firm established to exploit
rDNA technology
June 23, 1976 NIH released first guidelines for recombinant DNA experimentation
1977 human growth hormone genetically engineered
1977 methods for rapid DNA sequencing, discovery of split genes, and somatostanin by rDNA
Frederick Sanger developed rapid DNA sequencing techniques
1978 human insulin produced in E-coli
October 1978 Nobel prize given in recognition of discovery of restriction enzymes and their application to the problems of molecular genetics
December 1978 Biogen filed preliminary UK patent for technique to clone hepatitis B DNA and antigens

1979 insulin synthesized by using rDNA; first human viral antigen
1979 first DNA fragments of Epstein Barr virus cloned
February 1979 University of Edinburgh scientists published the successful isolation and cloning of DNA fragments of the hepatitis B virus in Escherichia coli
May 1979 - October 1979 Pasteur Institute scientists reported successful cloning of hepatitis B DNA in Escherichia coli
August 30, 1979 UCSF scientists announced the successful cloning and expression of HBsAG in Escherichia coli
December 21, 1979 Biogen applied for European patent to clone fragment of DNA displaying hepatitis B antigen specificity
1980 genetic engineering recognized for patenting; beginning of the creation of numerous biotech companies
1980 first patent awarded for gene cloning
1980 Cesar Milstein proposed the use of recombinant DNA to improve monoclonal antibodies
July 31, 1980 UCSF scientist published method to culture HBsAG antigens in cancer cells
September 1980 scientists reported the first successful development of transgenic mice
1981 foot and mouth disease viral antigen cloned
July 1981 UCSF and Merck filed patent to synthesize HBsAG in recombinant yeast
July 10, 1981 complete library of overlapping DNA fragments of Epstein Barr vvirus cloned
1982 first rDNA animal vaccine approved for sale in Europe (colibaccillosis) - first rDNA
pharmaceutical insulin approved for sale in USA and UK
first successful transfer of a gene from one animal species to another, a transgenic mouse
carrying the gene for rat growth hormone - first transgenic plant produced, using an agrobacterium transformation system
1982 commercial production of coli of genetically engineered human insulin, isolation, cloning and characterization of human cancer gene
March 9, 1982 Eli Lily company started
October 1982 first recombinant DNA drug approved

1983 first successful transfer of a plant gene from one species to another
1983 engineered Ti- plasmid used to transform plants
June 1, 1984 genetically engineered vaccine against hepatitis B reported to have positive trial results
Huntington’s disease is the first mapped genetic disease
1984 first chimeric monoclonal antibodies developed, laying foundation for safer and more effective monoclonal antibody therapeutics
1985 US patent office extends patent protection to genetically engineered plants
1985 insertion of cloned gene from salmonella into tobacco plant to make resistant to herbicide glyphosphate
1985 development of PCR technique
1986 transgenic pigs produced carrying the gene for human
growth hormone
1986 development of gene gun
May 1986 first humanized monoclonal antibody created
1986 first genetically engineered vaccine against hepatitis B approved
June 1986 interferon approved for treating hairy cell leukemia
December 1986 genetically engineered hepatitis B vaccine, Engerix-B, approved in Belgium
December 18, 1986 results released from first small-scale clinical trial of recombinant interferon-alpha therapy for post-transfusion chronic hepatitis B
1987 first field trials in USA of transgenic plants - tomatoes with
a gene for insect resistance
first field trials in USA of genetically engineered micro organism
1987 during the process of analyzing the E. coli genome for genes associated with phosphate metabolism, researchers made an unexpected discovery - they found 5 homologous sequences of 29 nucleotides arranged in direct repeats with 32 nucleotides as spacing in the end region of one gene - this repetitive sequence was later defined as CRISPR
1988 Campath-1H is created; the first clinically useful humanized monoclonal antibody
1988 US patent office extends patent protection to genetically engineered animals
first genetically modified organism approved
human genome mapping project initiated
April 12, 1988 OncoMouse patent granted
1989 first field test of genetically engineered virus (baculovirus) that kills cabbage looper caterpillars
January 1989 genetically engineered hepatitis B vaccine, GenHevac, approved in France
1990 the first gene found to be associated with increased susceptibility to familial breast and ovarian cancer is identified
1990 the human genome project begins
1991 production of first transgenic pigs and goats, manufacture of human hemoglobin
first test of gene therapy on human cancer patients
July 13, 1992 FDA approved the use of genetically engineered interferon-alpha, Intron A, for the treatment of hepatitis B
1993 Halophilic archaea are found to be able to adapt to environmental changes in salinity - while studying this, scientists discovered 30 base pair long DNA segments, repeated at regular distances
further sequencing experiments confirmed the region contained at these 14 near-perfectly conserved repeats flanked on end by a highly degenerated copy
October 13, 1993 Cetus Corporation was sold to Chiron and its patent rights sold for $300 million to Hoffman-LaRoche
December 30, 1993 FDA approved genetically engineered enzyme drug for cystic fibrosis
1994 the Flavr Savr tomato introduced; the first genetically engineered whole food approved for sale; fully human monoclonal antibodies produced in genetically engineered mice
March 8, 1994 National Institute of Health is created
December 22, 1994 first chimeric monoclonal antibody therapeutic approved for market
1995 haemophilus influenzae is the first bacterial genome sequenced
1996 Bermuda Principles established

1997 worlds first mammalian clone (Dolly) developed from a non-reproductive cell of an adult animal through cloning by nuclear transplantation
December 1997 first humanized monoclonal antibody approved
for market
1999 first human chromosome is decoded
2000 genetic code of the fruit fly is decoded
2002 four CRISPR-associated genes were identified in CRISPR-containing prokaryotes, each located adjacent to a CRISPR locus; these genes were absent in CRISPR-negative prokaryotes
2002 mouse is the first mammal to have its genome decoded
2003 the human genome project is completed
2005 three independent research groups demonstrated sequence similarity between spacer regions of CRISPRs and sequences of bacteriophages and plasmids; species containing CRISPR elements were protected from corresponding foreign invaders, demonstrating its function as an immune system
at the same time a pattern necessary for CRISPR to function as an immune system was first discovered in Streptococcus strains
later, other patterns were defined for organisms with different CRISPR systems
these short sequence motifs at joining protospacers were later termed PAM - protospacer adjacent motif
2007 the CRISPR/Cas system is demonstrated to function as an immune system in an experiment with a phage-host model system; a phagesensitive wild thermophilus strain and two virulent bacteriaophages were selected, and several phageresistant mutants were generated by challenging the wild-type strain with the phages; novel spacers derived from phage DNA were integrated into the CRISPR1 locus, conveying resistance to the corresponding phage - when the protospacer sequences were removed, resistance disappeared
2008 the molecular mechanisms underlying the CRISPR-based defense system had yet to be determined using protein tagging and affinity purification, a protein complex of five Cas proteins was identified the complex, Cascade (CRISP-associated complex for antiviral defense) was isolatd from E coli lysates; the CRISPR RNA endonuclease subunit of Cascade forms mature guide RNAs that are essential for antiviral defense

2012 in vitro studies demonstrated that the Cas9-crRNA complex functions as an RNA-guided endonuclease, with Cas9 causing DNA cleavage, and that crRNA base-paired to tracrRNA also directed Cas9 to introduce double-stranded breaks
2013 the ability to edit the genome of eukaryotic cells using CRISPR/Cas9 was demonstrated with targeted genome cleavage in human and mouse cells
CRISPR-on (nuclease-dead Cas9 protein) fused with a transcriptional activation domain and sgRNA is shown to effectively activate reporter genes in human and mouse cells; scientists were able to rescue mice with a dominant mutation in the Crygc gene that causes cataaracts, by injecting Cas9 mRNA and sgRNA into zygotes
2013 DNA Worldwide and Eurofins forensic discover identical twins have difference in their genetic makeup
2014 Cas9/sgRNA screens were established as a tool for systematic genetic analysis in mammalian cells
the CRISPR/Cas9 system has several potential benefits over existing functional screening methods, including the ability to study phenotypes that require a complete loss of function gene to be elicited, a low rate of false negatives in large-scale screens, and the lack of effect of off-target effects on screens; the genome engineering capabilities of CRISPR/Cas9 were demonstrated in primates, with successful targeting of several genes in monkey embryos by co-injection of Cas9 mRNA and sgRNAs
March 2014 promising results announced from trial conducted with HIV patients
2015 the first study demonstrating gene editing of human embryos was published; scientists used CRISPR/Cas9 to cleave the endogenous b-globin gene in tripronuclear zygotes
2016 scientists used CRISPR/Cas9 to improve chimeric antigen receptor (CAR) T-cell therapies by reducing alloreactivity
2019 a new gene editing system was characterized and engineered
2020 in early 2020, an outbreak of a novel respiratory virus, SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) rapidly evolved into a global pandemic; one of the greatest challenges was to develop a rapid, accurate testing method that could be used to identify infectious individuals and reduce disease transmission
a CRISPR based assay was developed for the detection and quantification of SARS-Co-2 RNA
unlike other assays, this approach enables direct detection of RNA without any additional manipulations such as amplification
a single guide RNA is used to detect viral RNA, and a flourescent signal may be measured with a mobile phone camera; as such, it is a potentially fast, accurate, and portable option for screening