Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors

Michael Gapinske, Jackson Winter, Devyani Swami, Lauren Gapinske, Wendy S. Woods, Shraddha Shirguppe, Angelo Miskalis, Anna Busza, Dana Joulani, Collin J. Kao, Kurt Kostan, Anne Bigot, Rashid Bashir, Pablo Perez-Pinera

Research output: Contribution to journalArticlepeer-review

Abstract

Duchenne muscular dystrophy is an X-linked monogenic disease caused by mutations in the dystrophin gene (DMD) characterized by progressive muscle weakness, leading to loss of ambulation and decreased life expectancy. Since the current standard of care for Duchenne muscular dystrophy is to merely treat symptoms, there is a dire need for treatment modalities that can correct the underlying genetic mutations. While several gene replacement therapies are being explored in clinical trials, one emerging approach that can directly correct mutations in genomic DNA is base editing. We have recently developed CRISPR-SKIP, a base editing strategy to induce permanent exon skipping by introducing C > T or A > G mutations at splice acceptors in genomic DNA, which can be used therapeutically to recover dystrophin expression when a genomic deletion leads to an out-of-frame DMD transcript. We now demonstrate that CRISPR-SKIP can be adapted to correct some forms of Duchenne muscular dystrophy by disrupting the splice acceptor in human DMD exon 45 with high efficiency, which enables open reading frame recovery and restoration of dystrophin expression. We also demonstrate that AAV-delivered split-intein base editors edit the splice acceptor of DMD exon 45 in cultured human cells and in vivo, highlighting the therapeutic potential of this strategy.

Original languageEnglish (US)
Pages (from-to)572-586
Number of pages15
JournalMolecular Therapy Nucleic Acids
Volume33
DOIs
StatePublished - Sep 12 2023

Keywords

  • AAV
  • CRISPR-Cas9
  • Duchenne muscular dystrophy
  • MT: RNA/DNA editing
  • adenine base editing
  • cytidine base editing
  • exon skipping
  • gene editing

ASJC Scopus subject areas

  • Drug Discovery
  • Molecular Medicine

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