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Through specific bioinformatic analysis, it was identified that Cas9 is composed of two regions: a gRNA binding recognition (REC) lobe with REC1 and REC2 subunits and a nuclease (NUC) lobe with a RuvC-like nuclease domain, His-Asn-His (HNH), a PAM-interacting (PI) domain, and a wedge (WED) domain [49]