Dual CRISPR-Cas3 system for inducing multi-exon skipping in DMD patient-derived iPSCs.

ABSTRACT: To restore dystrophin protein in various mutation patterns of Duchenne muscular dystrophy (DMD), the multi-exon skipping (MES) approach has been investigated. However, only limited techniques are available to induce a large deletion to cover the target exons spread over several hundred kilobases. Here, we utilized the CRISPR-Cas3 system for MES induction and showed that dual crRNAs could induce a large deletion at the dystrophin exon 45-55 region (∼340 kb), which can be applied to various types of DMD patients. We developed a two-color SSA-based reporter system for Cas3 to enrich the genome-edited cell population and demonstrated that MES induction restored dystrophin protein in DMD-iPSCs with three distinct mutations. Whole-genome sequencing and distance analysis detected no significant o

SUBMITTER: Kita Y 

PROVIDER: S-EPMC10545483 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature