TY  - JOUR
TI  - Multiplex Genome Engineering Using CRISPR/Cas Systems
AU  - Le Cong
AU  - F. Ann Ran
AU  - David Cox
AU  - Shuailiang Lin
AU  - Robert P. J. Barretto
AU  - Naomi Habib
AU  - Patrick D. Hsu
AU  - Xuebing Wu
AU  - Wenyan Jiang
AU  - Luciano A. Marraffini
AU  - Feng Zhang
PY  - 2013
JO  - Science
DO  - 10.1126/science.1231143
UR  - https://doi.org/10.1126/science.1231143
AB  - 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.
ER  -