Fig. 2: Screen of parameters that affect CRAFT-mediated RNA editing.

a Schematic guide tiling for 5′CRAFT rcRNAs along the target intron (top). Position and color of guide correlates with the quantification by flow cytometry presented as percent GFP positive cells and MFI (below) (Data are mean ± s.d. from n = 3 individual samples). b Lead guide candidate from the (a) (guide 4) was modified by extending the original 30 bp guide to 150 bp in 20bp step increments and evaluated in the splitGFP reporter assay. Quantitation by flow cytometry for the 5′CRAFT guide length: percent GFP positive cells (left) and MFI (right) (Data are mean ± s.d. from n = 3 individual samples). c The direct repeat in the rcRNA of the lead guide candidate from (a) was swapped with the direct repeat from alternative type VI CRISPR species and co-transfected with the cognate catalytically dead cas13 protein. Quantitation by flow cytometry is presented as percent GFP positive cells (left) and MFI (right) (Data are mean ± s.d. from n = 3 individual samples). d Schematic guide tiling for 3′CRAFT rcRNAs along the target intron (top). Position and color of guide correlates with the quantification by flow cytometry presented as percent GFP positive cells and MFI (below) (Data are mean ± s.d. from n = 3 individual samples). e Lead guide candidate from (d) (guide 2) was modified by extending the original 30 bp guide to 150 bp in 20 bp step increments and evaluated in the splitGFP reporter assay. Quantitation by flow cytometry for the 3′CRAFT guide length: percent GFP positive cells (left) and MFI (right) (Data are mean ± s.d. from n = 3 individual samples). f The direct repeat in the 3′rcRNA of the lead guide candidate of LMNA (guide 2) was swapped with the direct repeat from alternative type VI CRISPR species and co-transfected with the cognate catalytically dead cas13 protein. Quantitation by flow cytometry is presented as percent GFP positive cells (left) and MFI (right) (Data are mean ± s.d. from n = 3 individual samples). Statistical significance was determined by One-Way ANOVA with Tukey’s post-test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns not significant. Source data are provided as a Source Data file.