Tissue-specific and cell type-specific RNA interference in vivo

Manjeet K. Rao, Miles F. Wilkinson

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1494-1501
Number of pages8
JournalNature Protocols
Volume1
Issue number3
DOIs
StatePublished - Aug 2006
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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