Fig. 1: CRAFT efficiently rescues EGFP expression via precise RNA trans-splicing.

a Schematic of 5′ CRAFT RNA editing. b bright field (top) and fluorescent images (bottom) of HEK293 cells transfected with Psp-dCas13b, 5′ rcRNA, Psp-dCas13b and non-targeting 5′rcRNA, or Psp-dCas13b and a on-target 5′rcRNA. c Quantitation of flow cytometry for 5′CRAFT targeting LMNA intron showing percent GFP positive cells (left) and mean fluorescence intensity (MFI) (right) (Data are mean ± s.d. from n = 3 individual samples). d Schematic of 3′ CRAFT RNA editing. e bright field (top) and fluorescent images (bottom) of cells transfected with Rfx-dCas1d, 3′ rcRNA, Rfx-dCas13d and non-targeting 3′rcRNA, or Rfx-dCas13d and on-target 3′rcRNA. f Quantitation of flow cytometry for 3′CRAFT targeting LMNA intron showing percent GFP positive cells (left) and MFI (right) (Data are mean ± s.d. from n = 3 individual samples). g Schematic diagraming modified 5′rcRNAs (left) that lack the direct repeat (-DR), separate the spacer from the direct repeat (+stuffer), and original rcRNA. Quantitation by flow cytometry for of EGFP rescue using 5′rcRNA (Data are mean ± s.d. from n = 3 individual samples). h Quantification of EGFP rescue using orthogonal protein/RNA partners in 5′CRAFT; rcRNAs that contain the direct repeat from either PguCas13b (blue bar) or PspCas13b (green bar) along with either dPguCas13b enzyme (blue triangle) or dPspCas13b enzyme (green circle) (Data are mean ± s.d. from n = 3 individual samples). i Schematic diagraming modified 3′rcRNAs. Quantitation by flow cytometry for of EGFP rescue using 3′rcRNA (Data are mean ± s.d. from n = 3 individual samples). j Quantification of EGFP rescue using orthogonal protein/RNA partners in 5′CRAFT; rcRNAs that contain the direct repeat from either PguCas13b (blue bar) or PspCas13b (green bar) along with either LwaCas13d (blue bar) or RfxCas13d (green bar) along with either dLwaCas13d enzyme (blue triangle) or dRfxCas13d enzyme (green circle) (Data are mean ± s.d. from n = 3 individual samples). Statistical significance was determined by unpaired two-tailed Student t-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.