Figure 4.

Effects of guide-target mismatches on DNA and RNA cleavage by KmAgo. Reactions with guide DNA and target DNA (A), guide DNA and target RNA (B) and guide RNA and target DNA (C) were performed at 37°C for 15, 60 and 100 minutes, respectively, using 5′- P³²-labeled targets. Positions of the 5′-cleavage fragments (F) are indicated. In panels A and B, a series of different DNA guides contained substitutions at each guide position, shown above the panels. In panel C, a series of DNA targets contained substitutions at each position in the region of complementarity to the guide RNA, shown above the panel (see Supplementary Table S1 for full guide and target sequences). Means and standard deviations from three independent measurements.