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Beschreibung

CRISPR-associated endonuclease Cas12a (UniProt: A0A182DWE3, also known as CRISPR-associated endonuclease Cpf1, Lbcpf1) is a single crRNA-guided endonuclease of the class 2 CRISPR-Cas system that lacks tracrRNA and undergoes large conformational changes upon crRNA binding. Cpf1 has several features that can distinguish it from Cas9. Unlike Cas9 that uses dual-guide RNAs, Cpf1 is guided by a crRNA. It recognizes a T-rich PAM (T-rich protospacer adjacent motif), whereas Cas9 favors a G-rich PAM and Cpf1 generates staggered ends at its PAM-distal region, but Cas9 creates blunt ends at its PAM-proximal region. Cpf1 displays higher targeting specificity in mammalian cells when compared to Cas9. It has been shown that depending on the PAM sequences, LbCpf1 can undergo conformational changes to form altered interactions with the PAM-containing DNA duplexes, hence achieving the relaxed PAM recognition. (Ref.: Zetsche, B., et al. (2015). Cell 163(3), 759-77, Yamano, T., et al. (2017). Mol. Cell 67(4), 633-645, Gao, P., et al. (2016). Cell Res. 26(8), 901-913).