Quantitative analysis of conditional gene inactivation using rationally designed, tetracycline-controlled miRNAs.

ABSTRACT: The combination of RNA interference (RNAi) with the tetracycline-controlled transcription activation (tet) system promises to become a powerful method for conditional gene inactivation in cultured cells and in whole organisms. Here, we tested critical sequence elements that originated from miRNA mR-30 for optimal efficiency of RNAi-based gene knockdown in mammalian cells. Rationally designed miRNAs, expressed conditionally via the tet system, led to an efficient knockdown of the expression of both reporter genes and the endogenous mitotic spindle protein TPX2 in HeLa cells. Quantitative studies of the tet-controlled gene inactivation revealed that the residual expression of the target gene is an intrinsic attribute of all cells that cannot be eliminated either by increasing the miRNA to ta

SUBMITTER: Berger SM 

PROVIDER: S-EPMC2943624 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature