Microbial protein found in Streptococcus pyogenes M1 GAS
This article is missing information about homologs including other Cas9s, other Class 2 Type II CRISPR systems. Please expand the article to include this information. Further details may exist on the talk page.(September 2021)
CRISPR-associated endonuclease Cas9
S. pyogenes Cas9 in complex with sgRNA and its target DNA. PDB: 4OO8 [1]
Identifiers
Organism
Streptococcus pyogenes M1
Symbol
cas9
Alt. symbols
SpCas9
Entrez
901176
PDB
4OO8
RefSeq (mRNA)
NC_002737.2
RefSeq (Prot)
NP_269215.1
UniProt
Q99ZW2
Other data
EC number
3.1.-.-
Chromosome
Genomic: 0.85 - 0.86 Mb
Search for
Structures
Swiss-model
Domains
InterPro
Cas9
Identifiers
Symbol
?
InterPro
IPR028629
Cas9 (CRISPR associated protein 9, formerly called Cas5, Csn1, or Csx12) is a 160 kilodalton protein which plays a vital role in the immunological defense of certain bacteria against DNA viruses and plasmids, and is heavily utilized in genetic engineering applications. Its main function is to cut DNA and thereby alter a cell's genome. The CRISPR-Cas9 genome editing technique was a significant contributor to the Nobel Prize in Chemistry in 2020 being awarded to Emmanuelle Charpentier and Jennifer Doudna.[2]
More technically, Cas9 is a dual RNA-guided DNA endonuclease enzyme associated with the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) adaptive immune system in Streptococcus pyogenes.[3][4][5]S. pyogenes utilizes CRISPR to memorize and Cas9 to later interrogate and cleave foreign DNA, such as invading bacteriophage DNA or plasmid DNA.[4][6][7][8] Cas9 performs this interrogation by unwinding foreign DNA and checking for sites complementary to the 20 nucleotide spacer region of the guide RNA (gRNA). If the DNA substrate is complementary to the guide RNA, Cas9 cleaves the invading DNA. In this sense, the CRISPR-Cas9 mechanism has a number of parallels with the RNA interference (RNAi) mechanism in eukaryotes.
Apart from its original function in bacterial immunity, the Cas9 protein has been heavily utilized as a genome engineering tool to induce site-directed double-strand breaks in DNA. These breaks can lead to gene inactivation or the introduction of heterologous genes through non-homologous end joining and homologous recombination respectively in many laboratory model organisms. Research on the development of various cas9 variants has been a promising way of overcoming the limitation of the CRISPR-Cas9 genome editing. Some examples include Cas9 nickase (Cas9n), a variant that induces single-stranded breaks (SSBs) or variants recognizing different PAM sequences.[9] Alongside zinc finger nucleases and transcription activator-like effector nuclease (TALEN) proteins, Cas9 is becoming a prominent tool in the field of genome editing.
Cas9 has gained traction in recent years because it can cleave nearly any sequence complementary to the guide RNA.[4] Because the target specificity of Cas9 stems from the guide RNA:DNA complementarity and not modifications to the protein itself (like TALENs and zinc fingers), engineering Cas9 to target new DNA is straightforward.[10] Versions of Cas9 that bind but do not cleave cognate DNA can be used to locate transcriptional activator or repressors to specific DNA sequences in order to control transcriptional activation and repression.[11][12] Native Cas9 requires a guide RNA composed of two disparate RNAs that associate – the CRISPR RNA (crRNA), and the trans-activating crRNA (tracrRNA).[3] Cas9 targeting has been simplified through the engineering of a chimeric single guide RNA (chiRNA). Scientists have suggested that Cas9-based gene drives may be capable of editing the genomes of entire populations of organisms.[13] In 2015, Cas9 was used to modify the genome of human embryos for the first time.[14]
^Cite error: The named reference P24529477 was invoked but never defined (see the help page).
^"The Nobel Prize in Chemistry 2020". NobelPrize.org. Retrieved 2020-10-07.
^ abDeltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao Y, Pirzada ZA, Eckert MR, Vogel J, Charpentier E (March 2011). "CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III". Nature. 471 (7340): 602–607. Bibcode:2011Natur.471..602D. doi:10.1038/nature09886. PMC 3070239. PMID 21455174.
^ abcJinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (August 2012). "A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity". Science. 337 (6096): 816–21. Bibcode:2012Sci...337..816J. doi:10.1126/science.1225829. PMC 6286148. PMID 22745249.
^Oh HS, Diaz FM, Zhou C, Carpenter N, Knipe DM (2022-01-01). "CRISPR-Cas9 Expressed in Stably Transduced Cell Lines Promotes Recombination and Selects for Herpes Simplex Virus Recombinants". Current Research in Virological Science. 3: 100023. doi:10.1016/j.crviro.2022.100023. PMC 9629518. PMID 36330462.
^Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, et al. (March 2007). "CRISPR provides acquired resistance against viruses in prokaryotes". Science. 315 (5819): 1709–12. Bibcode:2007Sci...315.1709B. doi:10.1126/science.1138140. hdl:20.500.11794/38902. PMID 17379808. S2CID 3888761.
^Cite error: The named reference Garneau2010 was invoked but never defined (see the help page).
^Uddin, Fathema; M. Rudin, Charles; Sen, Triparna (August 2020). "CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future". Frontiers in Oncology. 10: 1387. doi:10.3389/fonc.2020.01387. PMC 7427626. PMID 32850447.
^Mali P, Esvelt KM, Church GM (October 2013). "Cas9 as a versatile tool for engineering biology". Nature Methods. 10 (10): 957–63. doi:10.1038/nmeth.2649. PMC 4051438. PMID 24076990.
^Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang L, Church GM (September 2013). "CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering". Nature Biotechnology. 31 (9): 833–8. doi:10.1038/nbt.2675. PMC 3818127. PMID 23907171.
^Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, Weissman JS, Qi LS (July 2013). "CRISPR-mediated modular RNA-guided regulation of transcription in eukaryotes". Cell. 154 (2): 442–51. doi:10.1016/j.cell.2013.06.044. PMC 3770145. PMID 23849981.
^Esvelt KM, Smidler AL, Catteruccia F, Church GM (July 2014). "Concerning RNA-guided gene drives for the alteration of wild populations". eLife. 3. doi:10.7554/eLife.03401. PMC 4117217. PMID 25035423.
^Cyranoski D, Reardon S (22 April 2015). "Chinese scientists genetically modify human embryos". Nature. doi:10.1038/nature.2015.17378. S2CID 87604469.
Cas9 (CRISPR associated protein 9, formerly called Cas5, Csn1, or Csx12) is a 160 kilodalton protein which plays a vital role in the immunological defense...
on a simplified version of the bacterial CRISPR-Cas9 antiviral defense system. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA)...
complementary to the CRISPR sequence. Cas9 enzymes together with CRISPR sequences form the basis of a technology known as CRISPR-Cas9 that can be used to edit genes...
RNA (sgRNA) is a short sequence of RNA that functions as a guide for the Cas9-endonuclease or other Cas-proteins that cut the double-stranded DNA and thereby...
CRISPR-associated nuclease such as Cas9 attaches to a tracrRNA–crRNA complex, which guides Cas9 to the invading protospacer sequence. But Cas9 will not cleave the protospacer...
modifications of DNA or RNA, similar to the better known CRISPR-Cas9 system. Cas12a is distinguished from Cas9 by a its single RuvC endonuclease active site, its 5'...
homology directed repair. The CRISPR/Cas9 system (CRISPR – Clustered Regularly Interspaced Short Palindromic Repeats, Cas9 – CRISPR-associated protein 9) is...
Francisco in early 2013. The technology uses a catalytically dead Cas9 (usually denoted as dCas9) protein that lacks endonuclease activity to regulate genes...
RNA-guided endonuclease (e.g., Cas9 or Cas12a) and its guide RNA, which can be easily altered to set the target. Cas9 is the most promising technology...
on Cas9 (from Type II systems), but other effectors like Cas12a (Type V) have been used as well. Cas9 Endonuclease Dead, also known as dead Cas9 or dCas9...
Doudna and Emmanuelle Charpentier were the first to propose that CRISPR-Cas9 (enzymes from bacteria that control microbial immunity) could be used for...
such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas9), are also improving the nutrition, taste and yield of crops. There are many...
contains regularly spaced CRISPR/Cas9 target sites into the genomes of progenitor cells. Alongside the barcode, Cas9 and sgRNA are introduced into the...
2015, a group of Chinese scientists used the gene-editing technique CRISPR/Cas9 to edit single-celled, non-viable embryos to see the effectiveness of this...
through a variety of methods, including homologous recombination, CRISPR-Cas9, and TALENs. One of the main advantages of gene knockouts is that they allow...
DNA site. It uses a fusion protein, consisting of a catalytically impaired Cas9 endonuclease fused to an engineered reverse transcriptase enzyme, and a prime...
marker for plant research to improve gene editing methods such as CRISPR/Cas9. In varying degrees most mammals have some skin areas without natural hair...
endless. For example, because it can be applied directly in the embryo, CRISPR/Cas9 reduces the time required to modify target genes compared to gene targeting...
invading viruses. The system, known as CRISPR-Cas9, consists of (a) an enzyme that destroys DNA (the nuclease Cas9) and (b) the DNA sequences of previously...
protein Cas9 to break the DNA backbones at specific target sequences. This system has shown a higher specificity than TALENs or ZFNs due to the Cas9 protein...
nations and increase accessibility to individual researchers. Likewise, CRISPR-Cas9 gene editing experiments can now be conceived and implemented by individuals...
repeats associated Cas9 (CRISPR/Cas9) are a new tool, further increasing the range of methods available. In particular CRISPR/Cas9 engineered endonucleases...
Jennifer Doudna and Emmanuelle Charpentier collaborated to develop the CRISPR/Cas9 system, a technique which can be used to easily and specifically alter the...