Ontology highlight
ABSTRACT:
SUBMITTER: Karvelis T
PROVIDER: S-EPMC4653880 | biostudies-literature | 2015
REPOSITORIES: biostudies-literature
Karvelis Tautvydas T Gasiunas Giedrius G Young Joshua J Bigelyte Greta G Silanskas Arunas A Cigan Mark M Siksnys Virginijus V
Genome biology 20151119
To expand the repertoire of Cas9s available for genome targeting, we present a new in vitro method for the simultaneous examination of guide RNA and protospacer adjacent motif (PAM) requirements. The method relies on the in vitro cleavage of plasmid libraries containing a randomized PAM as a function of Cas9-guide RNA complex concentration. Using this method, we accurately reproduce the canonical PAM preferences for Streptococcus pyogenes, Streptococcus thermophilus CRISPR3 (Sth3), and CRISPR1 ( ...[more]