2013
DOI: 10.1126/science.1231143
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Multiplex Genome Engineering Using CRISPR/Cas Systems

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 co… Show more

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Cited by 13,477 publications
(11,927 citation statements)
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References 36 publications
(32 reference statements)
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“…T7 Endonuclease I (T7E1) assay was performed to test the endogenous targeted cleavage ( Figure 4 a) because double‐stranded breaks in genomic DNA are mostly repaired by the indel‐forming nonhomologous end joining (NHEJ) pathway without template 29. LGCP resulted in efficient cleavage of the target gene ( Plk1 ) (26.2%) (Figure 4a).…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…T7 Endonuclease I (T7E1) assay was performed to test the endogenous targeted cleavage ( Figure 4 a) because double‐stranded breaks in genomic DNA are mostly repaired by the indel‐forming nonhomologous end joining (NHEJ) pathway without template 29. LGCP resulted in efficient cleavage of the target gene ( Plk1 ) (26.2%) (Figure 4a).…”
Section: Resultsmentioning
confidence: 99%
“…Compared with siRNA, which requires continuous administration to suppress certain genes, CRISPR/Cas9 enables genome‐level changes 29. Furthermore, the off‐target effect of siRNA remains a problem.…”
Section: Resultsmentioning
confidence: 99%
“…Since its innovation, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) system has been recognized as the most promising gene‐editing and gene‐regulation technique 2. The CRISPR/Cas9 system is an adaptive immunological response found in archaea and bacteria preventing invasion like viruses and plasmids 3.…”
Section: Introductionmentioning
confidence: 99%
“…CRISPR/Cas9 has attracted increasing attention since the report of its successful use in mammalian cells 36, 37 or human cells 37, 38. This approach has made gene editing easier and more efficient than the previous gene editing tools, such as zinc finger nuclease (ZFN) and transcription activator‐like effector nucleases (TALEN) 39.…”
Section: Discussionmentioning
confidence: 99%