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. 2020 Oct 16;11:5241. doi: 10.1038/s41467-020-19072-6

Fig. 2. The guide:target heteroduplex recognition.

Fig. 2

a The guide:target heteroduplex is located in the cleft between the REC and NUC lobes. b Magnified view of interactions between Cas12i2 and nucleotide (−1) of the target dsDNA. c Interaction of the WED domain with the RNA:DNA hybride at position +1. d Mutation of residues interacting with dsDNA or RNA:DNA duplex at position 1–2 impairs the DNA cleavage activity by Cas12i2. Linear plasmid DNA cleavage assay using the 300 ng linearized plasmid DNA as substrate in the presence of 200 nM mutants or wild-type Cas12i2–crRNA complex at 37 °C for 15 min. e Linear plasmid DNA cleavage assay showing that the length of the guide:target heteroduplex affects the cleavage activity of Cas12i2. f Linear plasmid DNA cleavage assay using single mismatched target DNA.