Fig. 2: Analysis of R-loop formation by dCas12a across different orthologs, target sequences, and mismatches.
From: Dynamic basis of supercoiling-dependent DNA interrogation by Cas12a via R-loop intermediates
A Left: histogram of measured change in equilibrium twist as a function of the imposed twist on the tether, aggregated from all recorded binding events for dAsCas12a with a FM sgRNA. The plot transitions from unwinding to rewinding at a twist of −5. Right: scatter plot of lifetime and change in equilibrium twist for all Steppi-scored states across all binding events, colored by twist density at state onset. Boxes highlight clusters of dwells at ~5 bp unwound ascribed to the seed region (black) and at >20 bp unwound ascribed to downstream unwinding (magenta). B–H same as (A) but with different gRNA, target sequence, and enzyme ortholog conditions. Boxes highlight clusters of dwells at ~20 bp ascribed to the full R-loop (red), ~17 bp ascribed to a distal intermediate (blue), and ~5 bp ascribed to the seed (black). See Supplementary Table 2 for trace statistics. Source data are provided for this figure.
