Genome-wide analysis reveals characteristics of off-target sites bound by the Cas9 endonuclease

By Cem Kuscu, Ş. Arslan, Ritambhara Singh, Jeremy Thorpe, Mazhar Adli. Nature Biotechnology, 2014

761 citations recorded by the source corpus.

View Paper: https://www.semanticscholar.org/paper/1be2c15562a08e310af77736b3f11e1cdb0e8d3d

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In Molecular Biology

Citations in the corpus

CRISPR/Cas9 in Genome Editing and Beyond. (2016, background): “However, it is necessary to carefully set up proper controls and compare its results with complementary experiments for validation, given that many studies have reported extensive genome-wide off-target dCas9-binding events (89, 90, 189, 227, 228).”
Target specificity of the CRISPR-Cas9 system (2014, neutral): “We and other groups recently generated the first maps of dCas9 binding on mammalian genomes [37,44,45]; all three studies revealed a large number of binding sites, for example up to six thousand in mouse embryonic stem cells, as well as substantial variation (200 fold) in the number of offtarget sit”
Highly Specific Epigenome Editing by CRISPR/Cas9 Repressors for Silencing of Distal Regulatory Elements (2015, neutral): “Previous reports have found evidence that dCas9 promiscuously binds the genome outside of the target site 29, 30, 41 .”
GUIDE-Seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases (2014, neutral): “…identify the seven off-target sites previously identified by ChIP-Seq and reported to be targets of mutagenesis by Cas9, we believe this is because those sites were incorrectly identified in that earlier study 18 as bona fide off-target cleavage sites (Supplementary Results and Supplementary Fig.”
In vivo genome editing using Staphylococcus aureus Cas9 (2015, background): “Although a number of studies have employed sequence similarity-based off-target search 22,26–30 or dCas9-ChIP 31,32 to predict off-target sites for Cas9, such approaches cannot assess the nuclease activity of Cas9 in a comprehensive and unbiased manner.”
Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation (2014, background): “Second, CRISPRi activity is highly sensitive to mismatches between the sgRNA and target DNA, suggesting that off-target binding of dCas9 observed in ChIP-seq experiments is too transient to impact transcription (Duan et al., 2014; Kuscu et al., 2014; Wu et al., 2014).”
The new frontier of genome engineering with CRISPR-Cas9 (2014, neutral): “Re-sults showed that in both human embryonic kidney (HEK293) cells ( 83 ) and mouse embryonic stem cells (mESCs) ( 84 ), a catalytically inactive version of Cas9 bound to many more sites than those matching the sequence of the sgRNA used in each case.”