| 초록 |
Three arm junction RNA nanostructures (Y-RNA) can provide dual roles of structural function and the biological function such as RNA interference (RNAi) by inserting siRNA sequences (anti-GFP, RFP, and BFP) into the each arm of Y-RNA. Through sequence and length optimization of the arm of Y-RNA, Y-RNA is carefully designed to be Dicer substrate for enhanced RNAi potency as well as prolonged effect. Interestingly, the Dicer processing and loading of the Y-RNA into the RNA Induced Silencing Complex (RISC) was highly dependent on the physical structure and sequence design of the three-arm junction. In addition, we enzymatically prepared Dicer substrate Y-RNA with a large scale using rolling circle transcription (RCT) and site-specific cleavage. Depend on cleavage site of helper, various Y-RNA can be produced from a single DNA template, inducing controlled RNAi effect. Our study can provide clarified RNAi nanostructures for programmable as well as simultaneous gene silencing in vitro. |
