RNA interference using a plasmid construct expressing short-hairpin RNA.

Methods in molecular biology (Clifton, N.J.) (2008-03-29)
Serene R Lai, Lucy G Andrews, Trygve O Tollefsbol

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.

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G 418 disulfate salt, powder, BioReagent, suitable for cell culture