Myospreader improves gene editing in skeletal muscle by myonuclear propagation

• 成果类型: Article
• 署名作者: Poukalov, Kiril K.; Valero, M. Carmen; Muscato, Derek R.; Adams, Leanne M.; Chun, Heejae; Lee, Young Il; Andrade, Nadja S.; Zeier, Zane; Sweeney, H. Lee; Wang, Eric T.
• 署名单位: State University System of Florida; University of Florida; State University System of Florida; University of Florida; State University System of Florida; University of Florida; State University System of Florida; University of Florida; State University System of Florida; University of Florida; University of Miami
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-8625
• DOI: 10.1073/pnas.2321438121
• 发表日期: 2024-05-07
• 关键词: muscular-dystrophy; messenger-rna; genomic dna; protein; mouse; transcription; localization; STABILITY; transport; sequence

摘要

Successful CRISPR/Cas9- based gene editing in skeletal muscle is dependent on efficient propagation of Cas9 to all myonuclei in the myofiber. However, nuclear- targeted gene therapy cargos are strongly restricted to their myonuclear domain of origin. By screening nuclear localization signals and nuclear export signals, we identify Myospreader, a combination of short peptide sequences that promotes myonuclear propagation. Appending Myospreader to Cas9 enhances protein stability and myonuclear propagation in myoblasts and myofibers. AAV- delivered Myospreader dCas9 better inhibits transcription of toxic RNA in a myotonic dystrophy mouse model. Furthermore, Myospreader Cas9 achieves higher rates of gene editing in CRISPR reporter and Duchenne muscular dystrophy mouse models. Myospreader reveals design principles relevant to all nuclear- targeted gene therapies and highlights the importance of the spatial dimension in therapeutic development.