Ribozyme-activated mRNA trans-ligation enables large gene delivery to treat muscular dystrophies

• 成果类型: Article
• 署名作者: Lindley, Sean R.; Subbaiah, Kadiam C. Venkata; Priyanka, Fnu; Poosala, Pornthida; Ma, Yijie; Jalinous, Leila; West, Jason A.; Richardson, William A.; Thomas, Tamlyn N.; Anderson, Douglas M.
• 署名单位: University of Rochester; University of Rochester; University of Rochester
• 刊物名称: SCIENCE
• ISSN/ISSBN: 0036-9761
• DOI: 10.1126/science.adp8179
• 发表日期: 2024-11-15
• 页码: 762-767
• 关键词: expression; dysferlin; generation; mechanism; myopathy; ligase; repair; rtcb; dna

摘要

Ribozymes are small catalytic RNA sequences capable of nucleotide-specific self-cleavage found widespread in nature. Ribozyme cleavage generates distinct 2 ',3 '-phosphate and 5 '-hydroxyl termini that resemble substrates for recently characterized RNA repair pathways in cells. We report that ribozyme cleavage of two separate mRNAs activated their scarless trans-ligation and translation into full-length protein in eukaryotic cells, a process that we named StitchR (for Stitch RNA). Optimization of StitchR activity in mammalian cells resulted in a similar to 900-fold increase in protein expression that approached levels observed for genes expressed from single vectors. We demonstrate that StitchR can be harnessed for effective dual adeno-associated virus gene therapies to correct muscular dystrophies by restoring large functional muscle proteins to endogenous levels in vivo.