Inhibition of gene function in mammalian cells using short-hairpin RNA (shRNA).
RNAi is now the preferred method for silencing gene expression in a variety of systems. In this chapter we describe the procedure for applying short-hairpin RNA (shRNA) to study gene function. Detailed descriptions of target site selection, shRNA construction, shRNA transfection and target knockdown validation are included.
Department of Pediatrics, University Hospital of North-Norway, Tromsø, Norway. joern.remi.henriksen@uit.no
Jørn Remi
JR
Henriksen
Jochen
J
Buechner
Cecilie
C
Løkke
Trond
T
Flægstad
Christer
C
Einvik
D000818:Animals; D020029:Base Pairing; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005796:Genes / Q000235:genetics / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018076:DNA, Complementary; D009841:Oligonucleotides; D034741:RNA, Small Interfering