Figure 1.

DNA cleavage by the TmaCmr complex is transcript-dependent. A, schematic depicting the interaction between the crRNA and the RNA protospacer. B, 5′-labeled ssDNA substrate was incubated with the TmaCmr complex, Mn²⁺ and either a complementary (c) or noncomplementary (n) RNA target with 10 nucleotide noncomplementary 5′ and 3′ flanks. The reaction products were analyzed by denaturing PAGE. M denotes markers for the substrate and expected product. C, quantification of DNA cleavage products by the TmaCmr complex over time upon activation with RNA targets containing various 3′ and 5′ flanks. RNA targets represented by green circles have a noncomplementary PFS, purple triangles represent targets without a PFS (dark purple includes a 5′ flank, light purple lacks a 5′ flank), and pink squares represent targets with an anti-tag PFS.