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. 2020 Jul 15;6(29):eabb4054. doi: 10.1126/sciadv.abb4054

Fig. 1. A growth-based PAM screen yielded alternative motifs for the S. pyogenes Cas9.

Fig. 1

(A) Genetic circuit for linking PAM sequences to growth on d-xylose in E. coli. We call the resulting screen PAM-SEARCH. gRNA, guide RNA. (B) Schematic of the growth-based screen. NGS, next-generation sequencing. (C) Output of the screen performed with SpyCas9. The data are displayed as a nucleotide depletion plot, PAM wheel (8), and motif plot. Each dot represents a distinct 5-nt sequence within the library. See data file S1 for an interactive version of the PAM wheel based on the Krona plot. Results from a biological replicate of the screen can be found in fig. S3 and data file S2.