Multiplex genome engineering using CRISPR/Cas systems.
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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. |
Year of Publication | 2013
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Journal | Science
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Volume | 339
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Issue | 6121
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Pages | 819-23
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Date Published | 2013 Feb 15
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ISSN | 1095-9203
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URL | |
DOI | 10.1126/science.1231143
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PubMed ID | 23287718
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PubMed Central ID | PMC3795411
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Grant list | R01-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
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