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Overview
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SpCas9 is a DNA endonuclease guided by crRNA and tracrRNA (or sgRNA alone), which is derived from S. pyogenes. SpCas9 specifically recognizes and cleaves dsDNA target in a PAM (NGG)-dependent manner. The cleavage site of Cas9 in the target sequence is 3 bps away from the PAM site. The HNH and RuvC domains of SpCas9 are responsible for cleaving the target and non-target strands of dsDNA, respectively. In Sp-dCas9, both H840 and D10 are mutated to A, so that its HNH and RuvC domain are inactivated and Sp-dCas9 has only the target DNA binding activity, but not the cleaving activity.
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