Document Detail

RNA-guided human genome engineering via Cas9.
MedLine Citation:
PMID:  23287722     Owner:  NLM     Status:  MEDLINE    
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.
Prashant Mali; Luhan Yang; Kevin M Esvelt; John Aach; Marc Guell; James E DiCarlo; Julie E Norville; George M Church
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2013-01-03
Journal Detail:
Title:  Science (New York, N.Y.)     Volume:  339     ISSN:  1095-9203     ISO Abbreviation:  Science     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-15     Completed Date:  2013-02-21     Revised Date:  2013-08-19    
Medline Journal Info:
Nlm Unique ID:  0404511     Medline TA:  Science     Country:  United States    
Other Details:
Languages:  eng     Pagination:  823-6     Citation Subset:  IM    
Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
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MeSH Terms
Caspase 9 / chemistry*,  genetics
Chromosomes, Human, Pair 19 / genetics
Codon / genetics
DNA Cleavage
Gene Targeting / methods*
Genetic Engineering / methods*
Genetic Loci
Genome, Human / genetics*
Induced Pluripotent Stem Cells
Inverted Repeat Sequences / genetics*
K562 Cells
RNA / chemistry*,  genetics
Grant Support
Reg. No./Substance:
0/Codon; 63231-63-0/RNA; EC 3.4.22.-/Caspase 9
Comment In:
Nat Rev Genet. 2013 Feb;14(2):80   [PMID:  23322222 ]
Nat Methods. 2013 Mar;10(3):189   [PMID:  23565557 ]
Science. 2013 Feb 15;339(6121):768-70   [PMID:  23413345 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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