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Science DOI:10.1126/science.1231143

Multiplex genome engineering using CRISPR/Cas systems.

Publication TypeJournal Article
Year of Publication2013
AuthorsCong, L, F Ran, A, Cox, D, Lin, S, Barretto, R, Habib, N, Hsu, PD, Wu, X, Jiang, W, Marraffini, LA, Zhang, F
JournalScience
Volume339
Issue6121
Pages819-23
Date Published2013 Feb 15
ISSN1095-9203
KeywordsAnimals, Base Sequence, CRISPR-Cas Systems, DNA, DNA Cleavage, Genetic Engineering, Genetic Loci, Genome, Humans, Inverted Repeat Sequences, Mice, Microarray Analysis, Molecular Sequence Data, Mutagenesis, Recombinational DNA Repair, RNA, Streptococcus pyogenes
Abstract

Functional elucidation of causal genetic variants and elements requires precise genome editing technologies. The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats)/Cas adaptive immune system has been shown to facilitate RNA-guided site-specific DNA cleavage. We engineered two different type II CRISPR/Cas systems and demonstrate that Cas9 nucleases can be directed by short RNAs to induce precise cleavage at endogenous genomic loci in human and mouse cells. Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair with minimal mutagenic activity. Lastly, multiple guide sequences can be encoded into a single CRISPR array to enable simultaneous editing of several sites within the mammalian genome, demonstrating easy programmability and wide applicability of the RNA-guided nuclease technology.

URLhttp://www.sciencemag.org/cgi/pmidlookup?view=long&pmid=23287718
DOI10.1126/science.1231143
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/23287718?dopt=Abstract

Alternate JournalScience
PubMed ID23287718
PubMed Central IDPMC3795411
Grant ListR01-CA133404 / CA / NCI NIH HHS / United States
T32 GM007753 / GM / NIGMS NIH HHS / United States
R01 CA133404 / CA / NCI NIH HHS / United States
DP1 MH100706 / MH / NIMH NIH HHS / United States
R01 GM034277 / GM / NIGMS NIH HHS / United States
R01 NS073124 / NS / NINDS NIH HHS / United States
DP2 AI104556 / AI / NIAID NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
R01-GM34277 / GM / NIGMS NIH HHS / United States
DP1MH100706 / DP / NCCDPHP CDC HHS / United States
DP2AI104556 / AI / NIAID NIH HHS / United States