A DNA vector-based RNAi technology to suppress gene expression in mammalian cells

TLDR

RNA interference, triggered by synthetic 21‑nt siRNAs, has become a powerful reverse‑genetic tool for silencing genes in plants, C. elegans, Drosophila, and mammalian cells. The study aims to create a DNA vector-based technology that generates siRNAs in vivo to inhibit endogenous gene expression. The approach uses DNA templates that are transcribed and processed into functional siRNAs within mammalian cells.

Abstract

Double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful reverse genetic tool to silence gene expression in multiple organisms including plants, Caenorhabditis elegans , and Drosophila . The discovery that synthetic double-stranded, 21-nt small interfering RNA triggers gene-specific silencing in mammalian cells has further expanded the utility of RNAi into mammalian systems. Here we report a technology that allows synthesis of small interfering RNAs from DNA templates in vivo to efficiently inhibit endogenous gene expression. Significantly, we were able to use this approach to demonstrate, in multiple cell lines, robust inhibition of several endogenous genes of diverse functions. These findings highlight the general utility of this DNA vector-based RNAi technology in suppressing gene expression in mammalian cells.