Zea mays cultivar:Hi-Type II Raw sequence reads

To expand the repertoire of Cas9s available for genome targeting a new in vitro method for simultaneous examination of guide RNA and PAM requirements is presented. 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, the canonical PAM preferences for Streptococcus pyogenes, Streptococcus thermophilus CRISPR3 (Sth3) and CRISPR1 (Sth1) are accurately reproduced. Additionally, PAM and sgRNA solutions for a novel Cas9 protein from Brevibacillus laterosporus are provided by the assay and are demonstrated to support functional activity in vitro and in plants.