RT Journal
A1 Agudelo, Daniel
A1 Carter, Sophie
A1 Velimirovic, Minja
A1 Duringer, Alexis
A1 Rivest, Jean-François
A1 Levesque, Sébastien
A1 Loehr, Jeremy
A1 Mouchiroud, Mathilde
A1 Cyr, Denis
A1 Waters, Paula J.
A1 Laplante, Mathieu
A1 Moineau, Sylvain
A1 Goulet, Adeline
A1 Doyon, Yannick
T1 Versatile and robust genome editing with Streptococcus thermophilus CRISPR1-Cas9
JF Genome Research 
JO Genome Research 
YR 2020 
FD January 01 
VO 30 
IS 1 
SP 107 
OP 117 
DO 10.1101/gr.255414.119 
UL http://genome.cshlp.org/content/30/1/107.abstract 
AB 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.