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

Nat Commun. 2024-04; 
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, Anna Cereseto
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CRISPR RNAs/Cas9 Proteins(INACTIVE) … sgRNA scaffolds, were obtained as synthetic fragments from either Genscript or Genewiz. … of the Cas9 genes was obtained from Genscript and cloned into an expression vector for in … Get A Quote

摘要

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 uncharacteriz... More

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