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FEBS Lett. 2004 Nov 19;577(3):376-80.

Development of a tightly regulated U6 promoter for shRNA expression.

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1
Cancer Research, Global Pharmaceutical Research and Development, AP10/L01, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064, USA.

Abstract

Short hairpin RNAs (shRNAs) have been used to achieve stable target knockdown in a variety of biological systems. Here, we report the development of a tightly regulated tetracycline-responsive human U6 promoter for shRNA expression. By engineering two copies of the tet operators flanking the TATA box of the human U6 promoter, we created a U6 promoter derivative (2O2) that exhibited much lower basal transcriptional activity compared with recently reported inducible pol III dependent promoters. As a consequence of its tighter regulation, the 2O2 system greatly improved the success rate in making inducible knockdown cell lines.

PMID:
15556613
DOI:
10.1016/j.febslet.2004.10.033
[Indexed for MEDLINE]
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