GSE211446
Deep sequencing data from PAM screen of Pa83 type IV-A CRISPR-Cas system
In this work the favorable PAM sequences for a type IV-A CRISPR-Cas system were determined Overall design: E. coli containing a recombinant type IV-A CRISPR-Cas immune system from P. aeruginosa were challenged against a plasmid library consisting of a complementary target flanked by a randomized protospacer-adjacent motif (PAM) sequence. E coli containing a nonfunctioning recombinant system without a CRISPR were also challenged with the PAM library as a control. Both the targeting and control experiments were done in biological triplicate, and the plasmid from the surviving cells was then purified and the target sequence extracted with PCR. Sequence counts for each PAM were then calculated from the resulting Illumina seq data, and a depletion score for each PAM was calculated.