Inhibition of gene function in mammalian cells using short-hairpin RNA (shRNA)

Jørn Remi Henriksen; Jochen Buechner; Cecilie Løkke; Trond Flægstad; Christer Einvik

doi:10.1007/978-1-59745-248-9_13

Inhibition of gene function in mammalian cells using short-hairpin RNA (shRNA)

Methods Mol Biol. 2011:703:189-204. doi: 10.1007/978-1-59745-248-9_13.

Authors

Jørn Remi Henriksen¹, Jochen Buechner, Cecilie Løkke, Trond Flægstad, Christer Einvik

Affiliation

¹ Department of Pediatrics, University Hospital of North-Norway, Tromsø, Norway. joern.remi.henriksen@uit.no

PMID: 21125491
DOI: 10.1007/978-1-59745-248-9_13

Abstract

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.

MeSH terms

Animals
Base Pairing
Base Sequence
Blotting, Western
Cloning, Molecular
DNA, Complementary / genetics
Genes / genetics
Genes / physiology*
Genetic Vectors / genetics
Molecular Sequence Data
Oligonucleotides / genetics
RNA Interference*
RNA, Small Interfering / genetics*
Reverse Transcriptase Polymerase Chain Reaction

Substances

DNA, Complementary
Oligonucleotides
RNA, Small Interfering