Fig. 5

Different Cas9 scaffolds increase the number of CRISPR-SKIP target exons. a, b RT-PCR analysis demonstrates that SpCas9-VQR-BE3 (a) and SaCas9-KKH-BE3 (b) can induce exon skipping of BRCA2 exon 26 and RELA exon 10, respectively. c, d Deep sequencing of genomic DNA revealed that targeted mutations (red) introduced by SpCas9-VQR-BE3 were found in 0.93% of reads at the BRCA2 exon 26 splice acceptor (c), while SaCas9-KKH-BE3 induced targeted mutations in 46.61% of reads at RELA exon 10 splice acceptor (d). Deep sequencing was performed in biological duplicates, and the results were combined. e, f Quantification of the rate of exon skipping of BRCA2 exon 26 (e) and RELA exon 10 (f) by deep sequencing of mature mRNA, which was amplified by RT-PCR. RNAseq was performed on biological duplicates and a single estimate of the proportion and confidence intervals were obtained (“Methods”)