Fig. 6: Engineering of Cas13bt3 for high-fidelity cleavage.
From: Structural basis for the activation of a compact CRISPR-Cas13 nuclease
a Two hydrophobic clusters around residue K645. Color codes are defined as in Fig. 1c. b Normalized on-target and collateral cleavage activities of mutated Cas13bt3. c Representative gel image of the collateral cleavage of WTCas13bt3, Y643A, and Y647A at different protein concentrations. RNA sizes (in nt) are indicated. b, c At least three times each experiment was repeated independently with similar results. d Normalized cleavage activity as shown in (c). e On-target and collateral cleavage of Cas13bt3 variants in HEK293T cells. Differential changes of normalized MFI were induced by dCas13bt3, WTCas13bt3, hfCas13bt3, Y647A, and Y643A in HEK293 cell lines. f Statistical data analysis for relative degradation efficiency by Cas13bt3 variants from (e). Normalized MFI, mean fluorescence intensity relative to the dCas13bt3 condition. d–f Data are presented as means ± SDs (n = 3). Source data are provided in the Source Data File.
