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Rivest, Jean-François.

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Rivest
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Jean-François.
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Centre de recherche, CHU de Québec, Université Laval
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  • Publication
    Accès libre
    Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9
    (Cold Spring Harbor, N.Y. Cold Spring Harbor Laboratory Press, 2020-01-30) Rivest, Jean-François.; Carter, Sophie; Laplante, Mathieu; Duringer, Alexis; Moineau, Sylvain; Lévesque, Sébastien.; Loehr, Jérémy; Velimirovic, Minja; Waters, Paula J.; Agudelo, Daniel; Goulet, Adeline
    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.