Fig. 1: Cryo-EM structure of the RdCas12n–sgRNA–target DNA ternary complex.
From: Mechanisms and engineering of a miniature type V-N CRISPR-Cas12 effector enzyme
a Unrooted phylogenetic tree from representative Cas12n nucleases. AcCas12n-like and RdCas12n-like clades are shown in blue and pink, respectively. Arrows indicate Cas12n genes experimentally characterized in this study (RdCas12n, AcCas12n, CgCas12n and MlCas12n). b The tested Cas12n nucleases for purification. RdCas12n was used for final cryo-EM analysis. c Diagram of the sgRNA and target DNA used for cryo-EM analysis. d The domain structure of RdCas12n. CTD, C-terminal domain. Residues 1–10 and 268–539 (the NUC lobe) were not included in the final model. e Cryo-EM density map of the RdCas12n–sgRNA–target DNA ternary complex structure 1 (PDB ID: 9J09). f The overall structure of the RdCas12n–sgRNA–target DNA ternary complex structure 1 (PDB ID: 9J09). Disordered regions are indicated as dotted lines. The NUC region of the protein exhibits a disordered structure, and the delineation of the NUC lobe is tentative, marking a predicted domain where the NUC lobe is likely to be situated.
