Cpf1 is a CRISPR−Cas endonuclease that, as a genome engineering tool, may have several advantages over Cas9, such as the production of staggered cuts with overhangs rather than blunt end cuts. Both Cpf1 and Cas9 use a CRISPR RNA (crRNA) to recognize target DNA, which is then cleaved using a RuvC nuclease domain. However, target cleavage by Cas9 additionally requires a HNH nuclease domain, whereas a second nuclease domain had not been identified for Cpf1. Yamano et al. and Dong et al. report crystal structures of Cpf1 that reveal a second putative nuclease domain, with a novel fold and no detectable homologues. Using mutational analysis, Yamano et al. show that this domain cleaves the target DNA at a distal site to produce the staggered cut. Remarkably, Fonfara et al. found that, in addition to DNase domains, Cpf1 has an RNase domain, which processes precursor transcripts into crRNAs. Thus, Cpf1 is an all-in-one machine for enacting CRISPR−Cas immunity.
References
Yamano, T. et al. Crystal structure of Cpf1 in complex with guide RNA and target DNA. Cell http://dx.doi.org/10.1016/j.cell.2016.04.003 (2016)
Dong, D. et al. The crystal structure of Cpf1 in complex with CRISPR RNA. Nature http://dx.doi.org/10.1038/nature17944 (2016)
Fonfara, I. et al. The CRISPR-associated DNA-cleaving enzyme Cpf1 also processes precursor CRISPR RNA. Nature http://dx.doi.org/10.1038/nature17945 (2016)
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Attar, N. How Cpf1 cuts its CRISPR targets. Nat Rev Microbiol 14, 335 (2016). https://doi.org/10.1038/nrmicro.2016.71
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DOI: https://doi.org/10.1038/nrmicro.2016.71
