Fig. 5: Effect of LdCsm3 mutations on the ssDNA cleavage and binding of LdCsm. | Cell Discovery

Fig. 5: Effect of LdCsm3 mutations on the ssDNA cleavage and binding of LdCsm.

From: Characterization of a novel type III CRISPR-Cas effector provides new insights into the allosteric activation and suppression of the Cas10 DNase

Fig. 5

a Target RNA cleavage of LdCsm3 mutated derivatives. Fifty nM of S1–46 RNA were incubated with 50 nM of LdCsm or the indicated mutant derivatives for 10 min, and the samples were analyzed by denaturing PAGE. Duplex: Duplex of crRNA and substrate. b RNA-activated ssDNA cleavage by effectors carrying one of the constructed LdCsm3 mutants. Reaction conditions were the same as in Fig. 4b. c ssDNA binding by effectors carrying each of the constructed LdCsm3 mutants. Reaction conditions were the same as in Fig. 4c. d Relative ssDNA binding between the wild-type LdCsm effector and its LdCsm3 mutated derivatives. The ssDNA activity of LdCsm in non-homologous RNA was used as the standard and set up as 1. Results shown are average of three independent assays; bars represent the mean standard deviation (±SD). The red arrow indicates the ssDNA-LdCsm complex.

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