Cas9 protein: Difference between revisions
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Cas9 ([[Crispr|CRISPR]] associated [[ | Cas9 ([[Crispr|CRISPR]] associated [[Protein|protein]] 9) is a family of [[Endonuclease|endonucleases]] that could cut [[DNA|DNA]] at specific sites. The Cas9 proteins are shown to only function as a complex with two types of [[RNA|RNA]]: CRISPR-RNA ([[CrRNA|crRNA]]) and trans-activating CRISPR-RNA ([[TracrRNA|tracrRNA]]).<ref>Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829</ref> Both RNAs are required to guide the Cas9 proteins to specific cleaving sites on double stranded DNA.<br> | ||
In prokaryotes such as [[ | In prokaryotes such as [[Streptococcus pyogenes|S. pyogenes]], the CRISPR-Cas9 system works as a defence mechanism to detect and cleave foreign DNAs that may come from [[Plasmid|plasmids]] or [[Bacteriophage|bacteriophages]].<ref>Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829</ref> CRISPR-Cas9 can be applied in [[Genetic engineering|genetic engineering]]. Cas9 is shown to be useful in bacterial gene manipulation.<ref>Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013 Mar;31(3):233-9. doi: 10.1038/nbt.2508</ref> In human cells, Cas9 can be used to [[Gene knockout|knockout]] or edit [[Gene|genes]] with the use of custom guide RNAs (gRNA).<ref>Mali P1, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, et al. RNA-guided human genome engineering via Cas9. Science. 2013 Feb 15;339(6121):823-6. doi: 10.1126/science.1232033</ref> Cas9 proteins can be engineered to have higher levels of specificity when they target desired DNA sequences.<ref>Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. Rationally engineered Cas9 nucleases with improved specificity. Science. 2016 Jan 1;351(6268):84-8. doi: 10.1126/science.aad5227</ref><br> | ||
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Revision as of 10:44, 4 December 2016
Cas9 (CRISPR associated protein 9) is a family of endonucleases that could cut DNA at specific sites. The Cas9 proteins are shown to only function as a complex with two types of RNA: CRISPR-RNA (crRNA) and trans-activating CRISPR-RNA (tracrRNA).[1] Both RNAs are required to guide the Cas9 proteins to specific cleaving sites on double stranded DNA.
In prokaryotes such as S. pyogenes, the CRISPR-Cas9 system works as a defence mechanism to detect and cleave foreign DNAs that may come from plasmids or bacteriophages.[2] CRISPR-Cas9 can be applied in genetic engineering. Cas9 is shown to be useful in bacterial gene manipulation.[3] In human cells, Cas9 can be used to knockout or edit genes with the use of custom guide RNAs (gRNA).[4] Cas9 proteins can be engineered to have higher levels of specificity when they target desired DNA sequences.[5]
References
- ↑ Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829
- ↑ Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829
- ↑ Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013 Mar;31(3):233-9. doi: 10.1038/nbt.2508
- ↑ Mali P1, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, et al. RNA-guided human genome engineering via Cas9. Science. 2013 Feb 15;339(6121):823-6. doi: 10.1126/science.1232033
- ↑ Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. Rationally engineered Cas9 nucleases with improved specificity. Science. 2016 Jan 1;351(6268):84-8. doi: 10.1126/science.aad5227