Multiple shRNA expressing vector enhances efficiency of gene silencing.
RNA interference (RNAi) is the process of sequence-specific gene silencing. However, RNAi efficiency still needs to be improved for effective inhibition of target genes. We have developed an effective strategy to express multiple shRNAs (small hairpin RNA) simultaneously using multiple RNA Polymerase III (Pol III) promoters in a single vector. Our data demonstrate that multiple shRNAs expressed from Pol III promoters have a synergistic effect in repressing the target gene. Silencing of endogenous cyclophilin A (CypA) or key HIV viral genes by multiple shRNAs results in significant inhibition of the target gene.
Song
Jun
J
Division of Oral Biology and Medicine, UCLA School of Dentistry, Los Angeles, CA 90095, USA. junsong@ucla.edu
Giang
An
A
Lu
Yingchun
Y
Pang
Shen
S
Chiu
Robert
R
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D021984:Cyclophilin A / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics*; D006678:HIV / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D012320:RNA Polymerase III / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics
D034741:RNA, Small Interfering; D014764:Viral Proteins; D012320:RNA Polymerase III; D021984:Cyclophilin A