用于短发夹RNA（shRNA）表达的严格调控U6启动子的开发。

Development of a tightly regulated U6 promoter for shRNA expression.

作者信息

Lin Xiaoyu, Yang Jianguo, Chen Jun, Gunasekera Angelo, Fesik Stephen W, Shen Yu

机构信息

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

出版信息

FEBS Lett. 2004 Nov 19;577(3):376-80. doi: 10.1016/j.febslet.2004.10.033.

DOI:10.1016/j.febslet.2004.10.033

PMID:15556613

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.

摘要

短发夹RNA（shRNAs）已被用于在多种生物系统中实现稳定的靶标敲低。在此，我们报告了一种用于shRNA表达的受严格调控的四环素响应性人U6启动子的开发。通过在人U6启动子的TATA框两侧设计两个四环素操纵子拷贝，我们创建了一个U6启动子衍生物（2O2），与最近报道的诱导型聚合酶III依赖性启动子相比，其基础转录活性要低得多。由于其调控更严格，2O2系统大大提高了构建诱导型敲低细胞系的成功率。