Enhanced CRISPR-Cas9 Correction of Duchenne Muscular Dystrophy in Mice by a Self-complementary AAV Delivery System

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2020 Mar 5

PMID 32128412

Citations 82

Authors

Yu Zhang

Hui Li

Yi-Li Min

Efrain Sanchez-Ortiz

Jian Huang

Alex A Mireault

John M Shelton

Jiwoong Kim

Pradeep P A Mammen

Rhonda Bassel-Duby

Eric N Olson

Affiliations

Soon will be listed here.

Abstract

Duchenne muscular dystrophy (DMD) is a lethal neuromuscular disease caused by mutations in the dystrophin gene (). Previously, we applied CRISPR-Cas9-mediated "single-cut" genome editing to correct diverse genetic mutations in animal models of DMD. However, high doses of adeno-associated virus (AAV) are required for efficient in vivo genome editing, posing challenges for clinical application. In this study, we packaged Cas9 nuclease in single-stranded AAV (ssAAV) and CRISPR single guide RNAs in self-complementary AAV (scAAV) and delivered this dual AAV system into a mouse model of DMD. The dose of scAAV required for efficient genome editing were at least 20-fold lower than with ssAAV. Mice receiving systemic treatment showed restoration of dystrophin expression and improved muscle contractility. These findings show that the efficiency of CRISPR-Cas9-mediated genome editing can be substantially improved by using the scAAV system. This represents an important advancement toward therapeutic translation of genome editing for DMD.

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