StitchR: Ribozyme-activated mRNA Trans-ligation and Translation

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). Overall design: StitchR 3.0 Nt-GFP and Ct-GFP AAV1-serotyped virus particles were generated and used to co-transduce HEK293T cells. Cells were harvested 48 hours post-transduction.

我们报道，在真核细胞中，两种分离的mRNA经核酶（ribozyme）切割后，可激活其无痕反式连接并翻译为全长蛋白质，我们将该过程命名为StitchR（取自stitch RNA之意）。实验设计概况：制备了腺相关病毒血清型1（AAV1）包装的、携载StitchR 3.0 N端绿色荧光蛋白（Nt-GFP）与C端绿色荧光蛋白（Ct-GFP）的病毒颗粒，并将其共同转导至HEK293T细胞；转导后48小时收集细胞。