Extended Data Fig. 8: Structural differences between SpyCas9-sgRNA-NTS_TS and SpyCas9-crRNA-tracrRNA-NTS_TS complexes.
From: Trans-nuclease activity of Cas9 activated by DNA or RNA target binding
a, Structures of SpyCas9-sgRNA-NTS_TS complexes with the 16-nt (top), 17-nt (middle first) and 18-nt (middle second) target strands in same orientation. (Bottom) The structure of SpyCas9-sgRNA-NTS_TS18 complex in catalytic state (PDB: 7Z4J). b, Structures of SpyCas9-crRNA-tracrRNA-NTS_TS complexes with the 16-nt (upper), 17-nt (middle) and 18-nt (bottom) target strands. c, SEC purification of SpyCas9-sgRNA-NTS_TS18 and SpyCas9-crRNA-tracrRNA-NTS_TS18 complexes. SEC assays were repeated three times independently with similar results. d, Structural comparison of the REC1 and HNH domains, as well as the sgRNA and target DNA in the checkpoint and catalytic state complexes. The checkpoint complex structure is shown in gray. In addition to the magenta core of the REC1 domain, the catalytic complex structure is colored according to a.
