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KMnO4 probing of mutant g8 protospacers bound by Cascade–g8–crRNA. (A) Probing of complexes formed on WT, PAM (CCG, G-1T), and indicated protospacer mutant templates. Positions of permanganate-sensitive thymines are indicated by dark red numbers on the right side of the gel. (B) A schematic model of R-loop formed upon the recognition of g8 protospacer target. Thymines sensitive to permanganate oxidation are shown in red (see also panel A).
