Fig. 5: Cas12a R-loop free energy landscapes and supercoiling-dependent cleavage.
From: Dynamic basis of supercoiling-dependent DNA interrogation by Cas12a via R-loop intermediates
A–D Cartoons of free energy landscapes (ΔG in units of kBT), depicted based on the results plotted in Figs. 3–4 (see Methods; dashed lines denote transitions that were not directly quantified under the twist condition shown). A Example of 8 MM effect on AsCas12a R-loop landscape. B Negative supercoiling tilts the 8 MM landscape, making open states accessible even in the presence of the mismatch. C Comparison between AsCas12a and LbCas12a. D Comparison between Seq 1 and Seq 2 with LbCas12a. E Example gels and quantification of cleavage assays for supercoiled and nicked plasmids incubated with WT Cas12a under all mismatch conditions. % cleavage of supercoiled (blue) and nicked (red) plasmids after 5 min of incubation with the indicated WT Cas12a complex is plotted as mean ± s.d. across three independent experiments. Source data are provided for this figure.
