High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity
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Cites background from "High-throughput profiling of off-ta..."
...The amount of Cas9 enzyme expressed in the cell is an important factor in tolerance to mismatches (138)....
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Cites background from "High-throughput profiling of off-ta..."
..., 2013), these studies collectively demonstrate that Cas9 tolerates mismatches throughout the guide sequence in a manner that is sensitive to the number, position, and distribution of the mismatches (Fu et al., 2013; Hsu et al., 2013; Mali et al., 2013b; Pattanayak et al., 2013)....
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...…2012; Cong et al., 2013), these studies collectively demonstrate that Cas9 tolerates mismatches throughout the guide sequence in a manner that is sensitive to the number, position, and distribution of the mismatches (Fu et al., 2013; Hsu et al., 2013; Mali et al., 2013b; Pattanayak et al., 2013)....
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...…appear to be better tolerated when Cas9 is present at high concentrations (Hsu 1270 Cell 157, June 5, 2014 ª2014 Elsevier Inc. et al., 2013; Pattanayak et al., 2013), leading to higher off-target activity; decreasing Cas9 concentration significantly improves the on- to off-target ratio at…...
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..., 2014), which correspond with the seed sequence identified through guide sequence mutation experiments (Jinek et al., 2012; Cong et al., 2013; Fu et al., 2013; Hsu et al., 2013; Pattanayak et al., 2013; Mali et al., 2013b)....
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...Streptococcus pyogenes Cas9 specificity has been extensively characterized by multiple groups using mismatched guide RNA libraries, in vitro selection, and reporter assays (Fu et al., 2013; Hsu et al., 2013; Mali et al., 2013b; Pattanayak et al., 2013)....
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References
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