title: RNA interference using a plasmid construct expressing short-hairpin RNA.
abstract: RNA interference (RNAi) is one of the most commonly used procedures for gene targeting in today's cutting edge technology and has great potential for use in clinical therapy. Using a plasmid construct that exogenously expresses short-hairpin RNAs (shRNAs) targeting a desired gene transcript not only helps to study the downstream effects of a gene product but also offers an alternative to viral vectors for gene therapy. Using a plasmid vector to knockdown a gene allows for long-term and permanent gene knockdown, without the need to generate knockout genotypes. Here, we detail the methodology for constructing a plasmid targeting the human telomerase reverse transcriptase (hTERT) gene through RNAi using the Ambion pSilencer system.
authors: Department of Biology, University of Alabama Birmingham, Birmingham, AL, USA.
authors: Serene R
authors: SR
authors: Lai
authors: Lucy G
authors: LG
authors: Andrews
authors: Trygve O
authors: TO
authors: Tollefsbol
mesh_terms: D002460:Cell Line; D002999:Clone Cells; D003001:Cloning, Molecular; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D009841:Oligonucleotides / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology*; D019098:Telomerase / Q000037:antagonists & inhibitors / Q000235:genetics; D014162:Transfection; D014170:Transformation, Genetic / Q000187:drug effects
chemical_list: D009841:Oligonucleotides; D034741:RNA, Small Interfering; C509186:TERT protein, human; D019098:Telomerase