研究人员报告了通过核酶切割两个独立的mRNA，激活了它们的无痕转移连接，并在真核细胞中翻译成全长蛋白，这一过程被命名为StitchR（Stitch RNA）。在哺乳动物细胞中优化StitchR活性导致蛋白质表达量提高了约900倍，接近从单一载体表达的基因的表达水平。
研究人员展示了StitchR可以被利用于有效的双腺相关病毒基因治疗，该工具可通过恢复大分子功能性肌肉蛋白至体内内源性水平来纠正肌营养不良症。
研究人员表示，核酶是能够进行核苷酸特异性自我切割的小型催化RNA序列，广泛存在于自然界中。核酶切割产生的2′,3′-磷酸和5′-羟基末端类似于细胞中最近鉴定的RNA修复途径的底物。
附：英文原文
Title: Ribozyme-activated mRNA trans-ligation enables large gene delivery to treat muscular dystrophies
Author: Sean R. Lindley, Kadiam C. Venkata Subbaiah, Fnu Priyanka, Pornthida Poosala, Yijie Ma, Leila Jalinous, Jason A. West, William A. Richardson, Tamlyn N. Thomas, Douglas M. Anderson
Issue&Volume: 2024-11-15
Abstract: 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 ~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.
DOI: adp8179
Source: https://www.science.org/doi/10.1126/science.adp8179