Multiplex Genome Engineering Using CRISPR/Cas Systems
Name
Zhang_Multiplex Genome.pdf
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287.82 KB
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Author(s)
Cong, Le
Ran, F. Ann
Cox, David Daniel
Lin, Shuailiang
Barretto, Robert
Habib, Naomi
Hsu, Patrick
Wu, Xuebing
Jiang, Wenyan
Marraffini, Luciano A.
Date Issued
February 2013
Journal
Science
Publisher
American Association for the Advancement of Science (AAAS)
Citation
Cong, Le, F. Ann Ran, David Cox, Shuailiang Lin, Robert Barretto, Naomi Habib, Patrick D. Hsu, Xuebing Wu, Wenyan Jiang, Luciano A. Marraffini, and Feng Zhang. "Multiplex Genome Engineering Using CRISPR/Cas Systems" Science 339:6121 (2013), pp. 819-823.
Version
Author's final manuscript
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.
MIT Department
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
McGovern Institute for Brain Research at MIT
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Creative Commons Attribution-Noncommercial-Share Alike
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DOI of Published Version
https://doi.org/10.1126/science.1231143
