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RNA-Guided Human Genome Engineering via Cas9

Published on Feb 15, 2013in Science41.04
· DOI :10.1126/science.1232033
Prashant Mali29
Estimated H-index: 29
(Harvard University),
Luhan Yang15
Estimated H-index: 15
(Harvard University)
+ 5 AuthorsGeorge M Church G M135
Estimated H-index: 135
(Wyss Institute for Biologically Inspired Engineering)
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.
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  • References (46)
  • Citations (4460)
References46
Newest
#1Le Cong (MIT: Massachusetts Institute of Technology)H-Index: 15
#2F. Ann Ran (MIT: Massachusetts Institute of Technology)H-Index: 9
Last.Luciano A. Marraffini (Rockefeller University)H-Index: 33
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#1Robert E. Thurman (UW: University of Washington)H-Index: 34
#2Eric Rynes (UW: University of Washington)H-Index: 8
Last.Benjamin Vernot (UW: University of Washington)H-Index: 19
view all 62 authors...
#1Martin Jinek (University of California, Berkeley)H-Index: 26
#2Krzysztof Chylinski (Umeå University)H-Index: 6
Last.Emmanuelle Charpentier (Umeå University)H-Index: 28
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#1Mina Rho (IU: Indiana University Bloomington)H-Index: 16
#2Yu Wei Wu (IU: Indiana University Bloomington)H-Index: 12
Last.Yuzhen Ye (IU: Indiana University Bloomington)H-Index: 29
view all 5 authors...
#1Blake Wiedenheft (HHMI: Howard Hughes Medical Institute)H-Index: 29
#2Samuel H. Sternberg (University of California, Berkeley)H-Index: 20
Last.Jennifer A. Doudna (LBNL: Lawrence Berkeley National Laboratory)H-Index: 92
view all 3 authors...
#1Neville E. Sanjana (McGovern Institute for Brain Research)H-Index: 21
#2Le Cong (McGovern Institute for Brain Research)H-Index: 15
Last.Feng Zhang (McGovern Institute for Brain Research)H-Index: 104
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#1Timothy R. Dreszer (UCSC: University of California, Santa Cruz)H-Index: 14
#2Donna Karolchik (UCSC: University of California, Santa Cruz)H-Index: 35
Last.Kate R. Rosenbloom (UCSC: University of California, Santa Cruz)H-Index: 25
view all 29 authors...
Cited By4460
Newest
#1Chang Chen (CAS: Chinese Academy of Sciences)
#2Jiajia Liu (CAS: Chinese Academy of Sciences)H-Index: 1
Last.Gang Liu (CAS: Chinese Academy of Sciences)H-Index: 16
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#1Fehad J. Khan (NIH: National Institutes of Health)H-Index: 1
#2Garmen Yuen (Johns Hopkins University)H-Index: 1
Last.Ji LuoH-Index: 28
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