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Enhancing SaCas9 Target Specificity

Enhancing SaCas9 Target Specificity

Enhancing SaCas9 target specificity by rational directed mutagenesis

The RNA-guided endonucleases of the CRISPR-Cas9 system, are becoming a robust genome editing tool in model organisms and hold immense promise for therapeutic applications. Specificity is the major concern of the CRISPR/Cas9 system as the Cas9 can cleave off-target sites that are not fully complementary to the guide sequence of the sgRNA. These specificity limitations would clearly complicate potential therapeutic applications and successful clinical use, thus off-target mutagenesis needs to be eliminated on genome wide scale. With the objective of limiting potential off-target without affecting on-target cleavage efficiencies, several approaches have been used to improve Cas9 specificity​...

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