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

Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5515-20. doi: 10.1073/pnas.082117599.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • Caenorhabditis elegans / genetics
  • Cell Line
  • DNA Modification Methylases / metabolism
  • Drosophila / genetics
  • Genetic Techniques*
  • Genetic Vectors*
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Models, Genetic
  • Plasmids / metabolism
  • RNA / metabolism*
  • RNA, Bacterial / genetics*
  • RNA, Bacterial / metabolism
  • Transfection
  • Tumor Cells, Cultured

Substances

  • RNA I
  • RNA, Bacterial
  • RNA
  • DNA Modification Methylases