Multiplex genome engineering using CRISPR/Cas systems.

Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F
Science. 2013 339 (6121): 819-23

PMID: 23287718 · PMCID: PMC3795411 · DOI:10.1126/science.1231143

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.

MeSH Terms (17)

Animals 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

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