RNA interference using a plasmid construct expressing short-hairpin RNA.
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.
Lai
Serene R
SR
Department of Biology, University of Alabama Birmingham, Birmingham, AL, USA.
Andrews
Lucy G
LG
Tollefsbol
Trygve O
TO
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
D009841:Oligonucleotides; D034741:RNA, Small Interfering; C509186:TERT protein, human; D019098:Telomerase