Figure 5.

Exonic in-frame deletion of a part of DMD exon 51 by NHBEJ editing

(A) Presentation of human DMD exon 51. Nucleotides and amino acids in reading frame, with five previous reported disease-causing mutations highlighted. (B) Schematic presentation of gRNA (pair 1: T1 + T2, pair 2: T3 + T4) targeting positions, length of deletions, and the resulting amino acid sequences after NHBEJ-mediated in-frame deletion. (C) PCR-genotyping results of HEK293T cells transfected with SpCas9 and either pair 1 (T1 + T2) or pair 2 (T3 + T4) gRNAs. Controls were transfected with SpCas9 only. (D) Semi-quantification of deletion efficiency in (C) by ImageJ. (E) Representative semi-estimation of NHBEJ efficiency based on the deletion products by Sanger sequencing and ICE analysis. (F) Boxplot of NHBEJ efficiency as mean and 1 SD (n = 3).