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Fig. 2 | Molecular Cancer

Fig. 2

From: Engineering the next-generation of CAR T-cells with CRISPR-Cas9 gene editing

Fig. 2

CRISPR/Cas9 components and Cas9 enzyme class variants. (Left) Endonuclease Cas9 can cleave a specific site within DNA as determined by the complementarity of the synthetic (crispr) crRNA to the target DNA strand, together with a (trans-activating) tracrRNA hairpin to provide structural stability. The crRNA and tracrRNA make up a singleĀ guide RNA (sgRNA) complex. Additional specificity is provided by the protospacer adjacent motif (PAM) that directs Cas9 to cut 3 base pairs upstream. a Cas9 cuts DNA via its HNH and RuvC domains that each cut a single DNA strand, resulting in a double-stranded break. b Mutations in one domain or both domains restrict(s) Cas9 to perform single-strand nicking (nCas9) or (c) to possess no catalytic activity (dCas9). Created with BioRender.com

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