RNAi expression vectors in mammalian cells.
RNA interference (RNAi) is a recently developed technique for gene silencing by introducing dsRNA into cells, and it is shown to work in mammalian cells when siRNAs are used. Several groups have developed vector-based siRNA expression systems that can induce RNAi in living cells. These vector systems use a pol III promoter, such as U6 or H1, and are classified into two groups based on the form of expressed RNAs: tandem-type and hairpin-type. Here, we describe how to generate these siRNA expression vectors and outline the experimental procedure for suppressing the expression of a reporter gene by transient transfection of a siRNA expression vector.
Miyagishi
Makoto
M
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Japan.
Taira
Kazunari
K
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008322:Mammals; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011401:Promoter Regions, Genetic / Q000235:genetics; D034622:RNA Interference / Q000502:physiology*; D016337:RNA, Catalytic / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D015183:Restriction Mapping; D014158:Transcription, Genetic
D017931:DNA Primers; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; C418481:hairpin ribozyme