Extended Data Fig. 6: Additional biochemical characterization of Cas7-11.

a, Schematic showing sequence of DiCas7-11 crRNA 1, targeting the ssRNA 1 target. b, Cleavage of ssRNA 1 target with crRNA 1 of varying DR and spacer lengths. c, Schematic showing sequence of DiCas7-11 crRNA 1, targeting the long MS2 target. d, Cleavage of long MS2 target with crRNA 1 with mutagenized DRs. e, DiCas7-11:crRNA complex binding to a complementary MS2 ssRNA target is determined by electrophoretic mobility shift assay (EMSA). EMSAs are performed for wild-type (WT) DiCas7-11, dead DiCas7-11, a non-targeting crRNA, and a MS2-targeting guide alone. f, DiCas7-11:crRNA complex binding to a complementary MS2 ssDNA target is determined by EMSA. g, Quantification of band intensities for non-targeting guide EMSA in e showing fitted curve (mean ± s.e.m.; n = 3). h, Quantification of band intensities for active DiCas7-11 with MS2 targeting guide EMSA in e showing fitted curve (mean ± s.e.m.; n = 3). i, Quantification of band intensities for MS2 targeting guide EMSA gel in e showing fitted curve (mean ± s.e.m.; n = 3). j, Quantification of band intensities for dead DiCas7-11 EMSA in e showing fitted curve (mean ± s.e.m.; n = 3). k, Quantification of band intensities for ssDNA EMSA gel in f showing fitted curve (mean ± s.e.m.; n = 3). l, RFP knockdown in E. coli is assayed via flow cytometry with wild-type and D429A/D654A DiCas7-11 protein expression. Data are mean ± s.e.m.; n = 3. m, Quantification of resistance conferred by top MS2-targeting DiCas7-11 spacers with active DiCas7-11, dead DiCas7-11, and no protein. Resistance is quantified as the highest surviving titer of MS2 phage that generates plaques (mean ± s.e.m.; n =3 ).