A novel approach for the construction of multiple shRNA expression vectors.
The application of RNA interference (RNAi) as a research and therapeutic tool depends on its ability to silence genes in a sequence-specific manner. Recent studies have reported that the effective knockdown of genes can be achieved by multiple short hairpin RNAs (shRNAs) in a single vector. Moreover, this approach can depress several genes simultaneously. However, current methods for the construction of multiple shRNA vectors often suffer from vector instability and are time-consuming. Here, we describe a simple, quick and low-cost approach to construct a single vector expressing four shRNA sequences driven by four different promoters. Using this vector, we were able to improve the gene silencing efficiency and make it possible to silence four different genes simultaneously, further expanding the application spectrum of RNAi, both in functional studies and therapeutic strategies.
Gou
Deming
D
Department of Physiological Sciences, Oklahoma State University, Stillwater, OK 74078, USA.
Weng
Tingting
T
Wang
Yang
Y
Wang
Zhixin
Z
Zhang
Honghao
H
Gao
Li
L
Chen
Zhongming
Z
Wang
Pengcheng
P
Liu
Lin
L
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D020868:Gene Silencing; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012320:RNA Polymerase III / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism*; D051381:Rats
D034741:RNA, Small Interfering; D012320:RNA Polymerase III