Table of Contents Author Guidelines Submit a Manuscript
Journal of Biomedicine and Biotechnology
Volume 2009, Article ID 514287, 7 pages
http://dx.doi.org/10.1155/2009/514287
Research Article

A Significant Increase of RNAi Efficiency in Human Cells by the CMV Enhancer with a tRNAlys Promoter

1The Key Laboratory of Chemical Biology, Division of Life Science, Graduate School at Shenzhen, Tsinghua University, Shenzhen, Guangdong 518055, China
2Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China
3Institute of Pharmacology, Medicine School, Tsinghua University, Beijing 100084, China

Received 19 November 2008; Revised 21 June 2009; Accepted 31 July 2009

Academic Editor: John McGregor

Copyright © 2009 Ma Weiwei et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

RNA interference (RNAi) is the process of mRNA degradation induced by double-stranded RNA in a sequence-specific manner. Different types of promoters, such as U6, H1, tRNA, and CMV, have been used to control the inhibitory effect of RNAi expression vectors. In the present study, we constructed two shRNA expression vectors, respectively, controlled by and CMV enhancer- promoters. Compared to the vectors with or U6 promoter, the vector with a CMV enhancer- promoter silenced pokemon more efficiently on both the mRNA and the protein levels. Meanwhile, the silencing of pokemon inhibited the proliferation of MCF7 cells, but the induction of apoptosis of MCF7 cells was not observed. We conclude that the CMV enhancer- promoter may be a powerful tool in driving intracellular expression of shRNA which can efficiently silence targeted gene.