Fig. 3: A flexible seed region at the 3′ end of the crRNA. | Nature Communications

Fig. 3: A flexible seed region at the 3′ end of the crRNA.

From: SCOPE enables type III CRISPR-Cas diagnostics using flexible targeting and stringent CARF ribonuclease activation

Fig. 3

a Schematic illustration of the endogenous TtCmr complex. b Different target RNAs with segments mismatches were used in an activity assay and analyzed on denaturing PAGE. c Impact of target RNAs with mismatches in the indicated segments on the production of cOA. d Schematic overview of the 46 nt crRNA complex (TtCmr-46). e Similar to panel b, using the 46 nt crRNA complex (TtCmr-46). f Similar to panel c, using 46 nt crRNA complex (TtCmr-46). g Schematic overview of the 40 nt crRNA complex (TtCmr-40). h Similar to panel b, using the 40 nt crRNA complex (TtCmr-40). i Similar to panel c, using the 40 nt crRNA complex (TtCmr-40). Target RNA contain a 5’ end Cy5 label (red circle); mismatched segments are indicated with S1–S6. Red triangles indicate the cleavage sites within TtCmr. CAR segments are indicated in green, seed segments are indicated in red. The results of the cleavage assays and cOA production assays displayed in this figure are representative results of three independent experiments (Supplementary Fig. S6). Error bars represent the standard deviation of the mean. Source data are provided as a Source data file.

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