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. Author manuscript; available in PMC: 2020 Dec 19.
Published in final edited form as: Mol Cell. 2019 Oct 24;76(6):938–952.e5. doi: 10.1016/j.molcel.2019.09.025

Figure 1. The overall structure of Nme1Cas9 in complex with truncated sgRNA.

Figure 1.

A. Domain organization of Nme1Cas9.

B. Schematic representation of the 135-nt sgRNA used for crystallization. The repeat:anti-repeat region is highlighted in green background. The stem-loops 1 and 2 are highlighted in pink and grey background, respectively. The black and blue dashed-line boxes indicate the truncated nts of the 122-nt and 102-nt sgRNAs, respectively. The cartoon diagram represents the state of Nme1Cas9 in this structure.

C. Crystal structure of the Nme1Cas9-sgRNA binary complex. Individual Nme1Cas9 domains are colored according to the scheme in Figure 1A.

D. DNA cleavage by wild-type and mutant Nme1Cas9 using linearized plasmid DNA containing a target sequence fully complementary to the sgRNA, and various sgRNAs containing different truncations.

E. Top: schematic representation of the partially double-stranded DNA target with its TS complementary to guide nts 17-24 (seed only). Bottom: crystal structure of the seed-only Nme1Cas9-sgRNA-dsDNA complex.

F. Conformational changes of Nme1Cas9 before (C) and after (E) PAM recognition and seed pairing.

See also Figures S1, S2, and Table S1.