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Multiple shRNAs expressed by an inducible pol II promoter can knock down the expression of multiple target genes

Authors :
Xu-Gang Xia
Hongxia Zhou
Zuoshang Xu
Source :
BioTechniques, Vol 41, Iss 1, Pp 64-68 (2006)
Publication Year :
2006
Publisher :
Taylor & Francis Group, 2006.

Abstract

RNA interference (RNAi) has been increasingly used for reverse genetics. Both pol III and pol II promoters have been used to synthesize short hairpin RNA (shRNA) for knockdown of gene expression in mammalian cells and animals. Compared with gene knockout approaches, RNAi has the advantage of being simple, quick, and low cost. Here we describe a method that enhances this advantage where knockdown of expression of multiple genes in the same cells is required. A tetracycline-regulated pol II promoter construct allows the expression of up to three shRNA genes that have been cloned into introns of a transcript bearing green fluorescent protein (GFP) coding sequences. This method may be used to establish stable knockdown cell lines and may also prove useful for investigating gene-gene interactions in transgenic animals.

Subjects

Subjects :
Biology (General)
QH301-705.5

Details

Language :
English
ISSN :
19409818 and 07366205
Volume :
41
Issue :
1
Database :
Directory of Open Access Journals
Journal :
BioTechniques
Publication Type :
Academic Journal
Accession number :
edsdoj.1ace0ee0c3e4f8395d6b964f42e5ecf
Document Type :
article
Full Text :
https://doi.org/10.2144/000112198