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CRISPR/Cas9 mechanism of action. The guide RNA (sgRNA) in association with the recognition (REC) lobe specifically hybridizes with the target dsDNA. Hybridization occurs through the spacer sequence, containing seed and non-seed regions. Cleavage of the dsDNA occurs only if PAM (5'-NGG-3') is presented, which is recognized by the nuclease (NUC) lobe. After recognition of the target dsDNA sequence and its PAM, the domains HNH and RuvC introduce a double-stranded break (DSB).
