Conditional RNA interference achieved by Oct-1 POU/rtTA fusion protein activator and a modified TRE-mouse U6 promoter.
RNA interference (RNAi) is a powerful technique and is widely used to down-regulate expression of specific genes in cultured cells and in vivo. In this paper, we report our development of a new tetracycline-inducible RNAi expression using a modified TRE-mouse U6 promoter in which the distal sequence element (DSE) was replaced by the tetracycline-responsive element (TRE). The modified TRE-mouse U6 promoter can be activated by a Tet-on version tetracycline-regulated artificial activator rTetOct which was constructed by fusing the rtTA DNA binding domain with the Oct-1 POU activation domain. This rTetOct/TRE-U6 system was successfully applied to conditionally and reversibly down-regulate the expression of endogenous p53 gene in MCF7 cells, and the expression of beta-defensin gene (mBin1b) either transiently expressed in COS7 cells or stably expressed in CHO cells.
Fei
Zhaoliang
Z
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Model Organism Research Center, SIBS, CAS, Shanghai, China.
Chen
Zheng
Z
Wang
Zhugang
Z
Fei
Jian
J
D000230:Adenocarcinoma / Q000378:metabolism; D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004318:Doxycycline / Q000494:pharmacology; D006801:Humans; D051379:Mice; D050811:Octamer Transcription Factor-1 / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034622:RNA Interference / Q000187:drug effects; D034741:RNA, Small Interfering / Q000502:physiology; D012342:RNA, Small Nuclear / Q000378:metabolism*; D013752:Tetracycline / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D023083:beta-Defensins / Q000096:biosynthesis
D050811:Octamer Transcription Factor-1; C494166:Pou2f1 protein, mouse; D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; C465595:Spag11 protein, mouse; D016159:Tumor Suppressor Protein p53; C405486:U6 small nuclear RNA; D023083:beta-Defensins; D013752:Tetracycline; D004318:Doxycycline