RNA-guided human genome engineering via Cas9.

Science 2013 Feb 3;339(6121):823-6. Epub 2013 Jan 3.

Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.

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.

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http://www.sciencemag.org/cgi/doi/10.1126/science.1232033
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February 2013
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References

(Supplied by CrossRef)

Wiedenheft et al.
Nature; Physical Science (London) 2012

Bhaya et al.
Annual review of genetics 2011

Terns et al.
Current opinion in microbiology 2011

Science 2012

PNAS 2012

Nucleic Acids Research 2011

Science 2002

Miyagishi et al.
Nature biotechnology 2002

Deltcheva et al.
Nature; Physical Science (London) 2011

Blood 2011

Sanjana et al.
2012

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