RNA-guided human genome engineering via Cas9
- PMID: 23287722
- PMCID: PMC3712628
- DOI: 10.1126/science.1232033
RNA-guided human genome engineering via Cas9
Abstract
Bacteria and archaea have evolved adaptive immune defenses, termed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems, that use short RNA to direct degradation of foreign nucleic acids. Here, we engineer the type II bacterial CRISPR system to function with custom guide RNA (gRNA) in human cells. For the endogenous AAVS1 locus, we obtained targeting rates of 10 to 25% in 293T cells, 13 to 8% in K562 cells, and 2 to 4% in induced pluripotent stem cells. We show that this process relies on CRISPR components; is sequence-specific; and, upon simultaneous introduction of multiple gRNAs, can effect multiplex editing of target loci. We also compute a genome-wide resource of ~190 K unique gRNAs targeting ~40.5% of human exons. Our results establish an RNA-guided editing tool for facile, robust, and multiplexable human genome engineering.
Figures
![Fig. 1](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/3712628/bin/nihms471334f1.gif)
![Fig. 2](https://cdn.statically.io/img/www.ncbi.nlm.nih.gov/pmc/articles/instance/3712628/bin/nihms471334f2.gif)
Comment in
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Technology: a CRISPR genome-editing tool.Nat Rev Genet. 2013 Feb;14(2):80. doi: 10.1038/nrg3409. Epub 2013 Jan 16. Nat Rev Genet. 2013. PMID: 23322222 No abstract available.
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Molecular biology. New tool for genome surgery.Science. 2013 Feb 15;339(6121):768-70. doi: 10.1126/science.1234726. Science. 2013. PMID: 23413345 No abstract available.
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RNA-guided gene editing.Nat Methods. 2013 Mar;10(3):189. doi: 10.1038/nmeth.2389. Nat Methods. 2013. PMID: 23565557 No abstract available.
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