Fig. 4: CRAFT is amenable to viral delivery and rewriting of endogenous mRNA.

a Schematic of stable HEK293 splitGFP reporter cell line generation accompanied by Sanger sequencing traces. b EGFP rescue in HEK293 cells that stably express the splitGFP reporter following transfection of 5′CRAFT plotted as percent GFP positive cells. c EGFP rescue in HEK293 cells that stably express the splitGFP reporter following transduction with 5′CRAFT (AAV genome schematics shown left) and plotted as percent GFP positive (right). d EGFP rescue in HEK293 cells that stably express the splitGFP reporter following transfection of 3′CRAFT plotted as percent GFP positive cells. e EGFP rescue in HEK293 cells that stably express the splitGFP reporter following transduction with 3′CRAFT (AAV genome schematics shown left) and plotted as percent GFP positive cells (right). f Schematic of cis- (left) trans- (right) endogenous LMNA transcripts highlighting a silent (C > G) snp in the trans-spliced. g Schematic of cis- (left) trans- (right) endogenous LMNA transcripts highlighting a silent (T > C) snp in the trans-spliced. h Sanger sequencing of the cis- (top) and trans- (bottom) spliced LMNA transcripts. i frequency of 5′CRAFT editing in endogenous LMNA transcripts. j Sanger sequencing of the cis- (top) and trans- (bottom) spliced LMNA transcripts. k frequency of 3′CRAFT editing in endogenous LMNA transcripts. l confocal microscopy images of N-terminal Flag tag introduced by 5′CRAFT Dapi (nuclear staining), AlexaFluor594 (Lamin A/C), and Flag (AlexaFluor488) (representative images from n = 2 independent experiments). m confocal microscopy images of C-terminal Flag tag introduced by 3′CRAFT (staining panel same as l). n volcano plot generated by DEseq2 analysis of differential gene expression from bulk RNA-seq in HEK293 cells transfected with dPspCas13b and either a targeting or non-targeting guide in the 5′ rcRNA. o volcano plot generated by DEseq2 analysis of differential gene expression from bulk RNA-seq in HEK293 cells transfected with dRfxCas13d and either a targeting or non-targeting guide in the 3′ rcRNA. All data are mean 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.