Fig. 1: In vitro RNase activity assays with the endogenous and reconstituted TtCmr complexes.

a Schematic illustration of the different reconstituted TtCmr complexes used in the activity assays shown in panel c, pre-loaded with either the 46 (TtCmr-46), 40 (TtCmr-40) or 34 nt (TtCmr-34) crRNA (top strand). Red triangles indicate the anticipated cleavage sites (α, β, γ and δ) in the 4.5 target RNA (bottom strand, Supplementary Table S1) by the endoribonuclease activity of the Cas7 subunits. The target RNA was radiolabeled at the 5′ end with ³²P γ-ATP (“P” in the red circle). b Denaturing PAGE analysis of the activity assay using a 5′ labeled target RNA complementary to the crRNA incubated with the endogenous TtCmr complex. A single stranded RNA marker (“M”) was used as size standards as indicated on the left. c Activity assays similar to panel b but using the reconstituted complexes. Discontinuous gel lanes are indicated by a dashed line. The results of the cleavage assays are representative results of three (b) or two (c) replicates (Supplementary Fig. 6). Source data are provided as a Source data file.