WB original data (Efficient exon skipping by base editor-mediated abrogation of exonic splicing enhancers)

Published: 27 September 2023| Version 1 | DOI: 10.17632/6wm6cszpfv.1
Han Qiu


Duchenne muscular dystrophy (DMD) is a severe genetic disease caused by the loss of the dystrophin protein. Exon skipping is a promising strategy to treat DMD by restoring truncated dystrophin. Here, we demonstrate that base editors (e.g., Targeted AID-mediated mutagenesis (TAM)) are able to efficiently induce exon skipping by disrupting functional redundant exonic splicing enhancers (ESEs). By developing an unbiased and high-throughput screening to interrogate exonic sequences, we successfully identify novel ESEs in DMD exons 51 and 53. TAM-CBE induces near-complete skipping of the respective exons by targeting these ESEs in patients’ iPSCs-derived cardiomyocytes. Combined with strategies to disrupt splice sites, we identify suitable sgRNAs with TAM-CBE to efficiently skip most DMD hotspot exons without substantial double-stranded breaks. Our study thus expands the repertoire of potential targets for CBE-mediated exon skipping in treating DMD and other RNA mis-splicing diseases.


Steps to reproduce

WT, N040006(patient), E50-N040006(patient & ΔE50), TAM-corrected-rep1 to TAM-corrected-rep2 (patient & ΔE50&51) cardiomyocytes which differentiated from iPSCs for 30 days were lysed and prepared protein samples. anti-Dystrophin (Sigma, D8168) and anti-mouse-IgG-HRP, Rabbit polyclonal to Cardiac Troponin T (Abcam, Ab45932), Rabbit monoclonal to Vinculin (Abcam, Ab129002) and anti-Rabbit-IgG-HRP were used to detect the target protein respectively. GEL Imaging System (AI680RGB) were used to detect the protein band, and analyzed by ImageJ.


Westlake University School of Life Sciences


Western Blot