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Journal Articles Genome Research Year : 2020

Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9

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.
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Dates and versions

hal-03050601 , version 1 (11-12-2020)

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Attribution - NonCommercial

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Daniel Agudelo, Sophie Carter, Minja Velimirovic, Alexis Duringer, Jean-François Rivest, et al.. Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9. Genome Research, 2020, 30 (1), pp.107-117. ⟨10.1101/gr.255414.119⟩. ⟨hal-03050601⟩
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