Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9

Authors: Agudelo, DanielCarter, SophieVelimirovic, Minja; Duringer, Alexis; Rivest, Jean-François; Levesque, SébastienLoehr, Jérémy; Waters, Paula J.; Laplante, MathieuMoineau, Sylvain; Goulet, Adeline
Abstract: Targeting definite genomic locations using CRISPR-Cas systems requires a set of enzymes with unique protospacer adjacent motif (PAM) compatibilities. To expand this repertoire, we engineered nucleases, cytosine base editors, and adenine base editors from the archetypal Streptococcus thermophilus CRISPR1-Cas9 (St1Cas9) system. We found that St1Cas9 strain variants enable targeting to five distinct A-rich PAMs and provide a structural basis for their specificities. The small size of this ortholog enables expression of the holoenzyme from a single adeno-associated viral vector for in vivo editing applications. Delivery of St1Cas9 to the neonatal liver efficiently rewired metabolic pathways, leading to phenotypic rescue in a mouse model of hereditary tyrosinemia. These robust enzymes expand and complement current editing platforms available for tailoring mammalian genomes.
Document Type: Article de recherche
Issue Date: 30 January 2020
Open Access Date: 31 March 2020
Document version: AM
This document was published in: Genome research, Vol. 30 (1), 107-117 (2020)
Cold Spring Harbor, N.Y. Cold Spring Harbor Laboratory Press
Alternative version: 10.1101/gr.255414.119
Collection:Articles publiés dans des revues avec comité de lecture

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