Journal of RNAi and Gene Silencing (August 2005), 1(1), 38-43

New Methods and Technologies

Design and application of a versatile expression vector for RNAi in mammalian cells

Xia Lin * and Xin-Hua Feng

ABSTRACT

Here we report a versatile mammalian expression vector called pRIGHT11 for production of small interfering RNA (siRNA) in cells. This vector uses opposing eukaryotic RNA polymerase III promoters U6 and H1 to drive the expression of short siRNA. We have demonstrated the effectiveness of pRIGHT11-generated siRNA in sequence-specific inhibition of transfected reporter genes and endogenous genes. Furthermore, this retrovirus-based vector can carry a random library of siRNA, and thus can be applied to rapid screening of novel genes involved in specific cellular responses.

KEYWORDS: RNAi, retroviral, polymerase III, Smad, vector, delivery, siRNA, shRNA

Michael E. DeBakey Department of Surgery, and Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA .

*Correspondence to: Xia Lin, Email: xialin@bcm.tmc.edu, Tel: +713 798 4899, Fax: +713 798 4093

© Copyright Lin and Feng

(Received 19 May 2005; Revised 16 June 2005; Accepted 16 June 2005)