Single small-interfering RNA expression vector for silencing multiple transforming growth factor-beta pathway components.

Amarsanaa Jazag; Fumihiko Kanai; Hideaki Ijichi; Keisuke Tateishi; Tsuneo Ikenoue; Yasuo Tanaka; Miki Ohta; Jun Imamura; Bayasi Guleng; Yoshinari Asaoka; Takao Kawabe; Makoto Miyagishi; Kazunari Taira; Masao Omata

doi:10.1093/nar/gni130

PubMed · 16113239

Single small-interfering RNA expression vector for silencing multiple transforming growth factor-beta pathway components.

Amarsanaa Jazag·Fumihiko Kanai·Hideaki Ijichi·Keisuke Tateishi·Tsuneo Ikenoue·Yasuo Tanaka·Miki Ohta·Jun Imamura·Bayasi Guleng·Yoshinari Asaoka·Takao Kawabe·Makoto Miyagishi·Kazunari Taira·Masao Omata

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Abstract

Although RNA interference (RNAi) is a popular technique, no method for simultaneous silencing of multiple targets by small-hairpin RNA (shRNA)-expressing RNAi vectors has yet been established. Although gene silencing can be achieved by synthetic small-interfering RNA (siRNA) duplexes, the approach is transient and largely dependent on the transfection efficiency of the host cell. We offer a solution: a simple, restriction enzyme-generated stable RNAi technique that can efficiently silence multiple targets with a single RNAi vector and a single selection marker. In this study, we succeeded in simultaneous stable knockdown of transforming growth factor beta (TGF-beta) pathway-related Smads--Smad2, Smad3 and Smad4--at the cellular level. We observed distinct phenotypic changes in TGF-beta-dependent cellular functions such as invasion, wound healing and apoptosis. This method is best suited for an analysis of complex signal transduction pathways in which silencing of a single gene cannot account for the whole process.

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Cell Line DNA-Binding Proteins Gene Expression Genetic Vectors Humans Phenotype Protein Serine-Threonine Kinases RNA Interference RNA, Small Interfering Receptor, Transforming Growth Factor-beta Type II Receptors, Transforming Growth Factor beta Signal Transduction Smad2 Protein Smad3 Protein Smad4 Protein Trans-Activators Transforming Growth Factor beta Journal Article Research Support, Non-U.S. Gov't

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Amarsanaa Jazag, Fumihiko Kanai, Hideaki Ijichi, Keisuke Tateishi, Tsuneo Ikenoue, Yasuo Tanaka, Miki Ohta, Jun Imamura, Bayasi Guleng, Yoshinari Asaoka, Takao Kawabe, Makoto Miyagishi, Kazunari Taira, Masao Omata. 2005-08-19. Single small-interfering RNA expression vector for silencing multiple transforming growth factor-beta pathway components.. https://doi.org/10.1093/nar%2Fgni130

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