Premium
Development of a tightly regulated U6 promoter for shRNA expression
Author(s) -
Lin Xiaoyu,
Yang Jianguo,
Chen Jun,
Gunasekera Angelo,
Fesik Stephen W.,
Shen Yu
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.10.033
Subject(s) - gene knockdown , small hairpin rna , promoter , tata box , rna interference , microbiology and biotechnology , promoter activity , rna polymerase iii , biology , chemistry , rna , gene expression , cell culture , genetics , gene , rna polymerase
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.