Tissue-specific and Cell Type-Specific RNA Interference in Vivo

Nat Protoc. 2006;1(3):1494-501. doi: 10.1038/nprot.2006.260.

Abstract

RNA interference (RNAi) is an efficient method for silencing genes in cultured cells. Here we describe a simple RNAi approach for silencing genes in a cell type-specific and tissue-specific way in vivo. The approach, which mimics the means by which naturally occurring 'microRNA's are generated, uses a tissue-specific polymerase II promoter to drive the expression of a short hairpin RNA (shRNA) directed against the gene target. The shRNA is cleaved by ubiquitously expressed endonucleases to form an active small interfering RNA of about 22 nt. As a proof of principle, it has been shown that expression of a shRNA directed against the transcription factor Wilms tumor 1 in transgenic mice reduces that protein specifically in nurse cells in the testis. Our transgenic RNAi approach offers a cost-effective means of rapidly (within months) addressing the function(s) of genes of interest in a wide variety of specific cell types and tissues in mice in vivo.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • DNA Polymerase II
  • Genes, Wilms Tumor
  • Genetic Techniques*
  • Genetic Vectors
  • Mice
  • RNA Interference*
  • RNA, Small Interfering / genetics*

Substances

  • RNA, Small Interfering
  • DNA Polymerase II