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.

中文翻译：

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核酶激活的 mRNA 反式连接使大基因递送能够治疗肌肉萎缩症


核酶是能够在自然界中广泛发现的核苷酸特异性自切割的小催化 RNA 序列。核酶切割产生不同的 2'，3'-磷酸和 5'-羟基末端，类似于细胞中最近表征的 RNA 修复途径的底物。我们报道了两个独立 mRNA 的核酶切割激活了它们在真核细胞中的无疤痕转连接和翻译成全长蛋白质，我们将这一过程命名为 StitchR（缝合 RNA）。优化哺乳动物细胞中的 StitchR 活性导致蛋白质表达增加 ~900 倍，接近从单个载体表达的基因观察到的水平。我们证明 StitchR 可用于有效的双重腺相关病毒基因疗法，通过在体内将大型功能性肌肉蛋白恢复到内源性水平来纠正肌营养不良。