Supplementary Figure 10: Expanded screening of PAM specificity for three Cascade homolog variants.
From: Harnessing type I CRISPR–Cas systems for genome engineering in human cells
a, FokI-PseCascade (with 17-aa FokI-Cas8 linker) editing efficiency as a function of PAM sequence. Target sites contained a fixed ATG PAM and a variable PAM at the second half-site, as shown on the x-axis. Each dot represents a single target site in HEK293 cells and n=6 (ATG), 8 (AAC, GAG, ATA, AAT), 9 (AGG), 12 (AAG), 14 (AAA) unique target sites, bar graph and error bars report mean and s.d. across sites, data with dual-AAG/ATG PAMs were identical to those shown in Fig. 3d. b, FokI-EcoCascade (with 17-aa FokI-Cas8 linker) editing as a function of PAM sequence. Target sites contained a fixed AAG PAM and a variable PAM at the second half-site, as shown on the x-axis. Each dot represents a single target site in HEK293 cells and n=6 (AAG), 8, (ATA), 9 (GAG, AAC), 10 (CGC), 11 (CCG), 12 (ATG), 13 (AGG), 15 (AAA) unique target sites, bar graph and error bars report mean and s.d. across sites. c, Same as in b, but with one PAM fixed as an ATG, n=6 (ATG), 8 (AAC, GAG, AAT, ATA), 9 (AGG), 12 (AAG), 14 (AAA) unique target sites, bar graph and error bars report mean and s.d. across sites, data with dual-AAG/ATG PAMs were identical to those shown in b. d, FokI-SthCascade (with 17-aa FokI-Cas8 linker) editing as a function of PAM sequence. Target sites contained a fixed GAA PAM and a variable PAM at the second half-site, as shown on the x-axis. Each dot represents a single target site in HEK293 cells and n=18 (TA), 28 (CC), 30 (AA), 31 (GA), 33 (CA) unique target sites, bar graph and error bars report mean and s.d. across sites.
