Fig. 5: Proximal mismatches limit Cas13d cleavage activity.
From: Massively parallel profiling of RNA-targeting CRISPR-Cas13d
A Schematic of the collateral cleavage assay. B Fluorescent cleavage time courses for matched target and nine representative mismatched target RNAs. Blue lines are distal mismatched sequences (positions 12-22). Orange lines are proximal mismatched sequences (positions 1-11). C The initial slope of 19 mismatched sequences. Slopes are calculated by the fluorescence changes during the first 20 minutes of the cleavage reaction and normalized to the matched target. Data are shown as mean and S.D. from two replicates. D Correlation of the cleavage slope with binding affinity (∆ABA). A subset of target RNAs retain strong binding but are cleavage-inactive (boxed region). Data are shown in mean ± S.D. from bootstrap analysis (y-axis). All sequence counts are detailed in the Source Data. E Schematic of mismatch-defined differentiation between SARS-CoV-2 variants of concern (VOC). F Fluorescent cleavage time courses for SARS-CoV-2 Wuhan and Delta VOCs. G The initial slope of the trace in (F). Slopes are calculated by the fluorescence changes during the first 20 minutes of the cleavage reaction. Data are shown as mean and S.D. from three replicates.
