CoCas9 is a compact nuclease from the human microbiome for efficient and precise genome editing.

Pedrazzoli, Eleonora; Demozzi, Michele; Visentin, Elisabetta; Ciciani, Matteo; Bonuzzi, Ilaria; Pezzè, Laura; Lucchetta, Lorenzo; Maule, Giulia et al. · Nat Commun · 2024

basic_science · Level V

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Abstract

The expansion of the CRISPR-Cas toolbox is highly needed to accelerate the development of therapies for genetic diseases. Here, through the interrogation of a massively expanded repository of metagenome-assembled genomes, mostly from human microbiomes, we uncover a large variety (n = 17,173) of type II CRISPR-Cas loci. Among these we identify CoCas9, a strongly active and high-fidelity nuclease with reduced molecular size (1004 amino acids) isolated from an uncultivated Collinsella species. CoCas9 is efficiently co-delivered with its sgRNA through adeno associated viral (AAV) vectors, obtaining efficient in vivo editing in the mouse retina. With this study we uncover a collection of previously uncharacterized Cas9 nucleases, including CoCas9, which enriches the genome editing toolbox.

Medical subject headings