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

Eleonora Pedrazzoli, Michele Demozzi, Elisabetta Visentin, Matteo Ciciani, Ilaria Bonuzzi, Laura Pezzè, Lorenzo Lucchetta, Giulia Maule, Simone Amistadi, Federica Esposito, Mariangela Lupo, Annarita Miccio, Alberto Auricchio, Antonio Casini, Nicola Segata () and Anna Cereseto ()
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Eleonora Pedrazzoli: University of Trento
Michele Demozzi: University of Trento
Elisabetta Visentin: University of Trento
Matteo Ciciani: University of Trento
Ilaria Bonuzzi: University of Trento
Laura Pezzè: Alia Therapeutics
Lorenzo Lucchetta: University of Trento
Giulia Maule: University of Trento
Simone Amistadi: University of Trento
Federica Esposito: Telethon Institute of Genetics and Medicine (TIGEM)
Mariangela Lupo: Telethon Institute of Genetics and Medicine (TIGEM)
Annarita Miccio: INSERM
Alberto Auricchio: Telethon Institute of Genetics and Medicine (TIGEM)
Antonio Casini: Alia Therapeutics
Nicola Segata: University of Trento
Anna Cereseto: University of Trento

Nature Communications, 2024, vol. 15, issue 1, 1-12

Abstract: 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.

Date: 2024
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DOI: 10.1038/s41467-024-47800-9

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