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RNAi in Cultured Mammalian Cells Using Synthetic SiRNAs

Overview
Journal Cold Spring Harb Protoc
Specialties Biomedical Engineering
Pathology
Date 2012 Sep 6
PMID 22949722
Citations 10
Authors
Kenneth Chang
Krista Marran
Amy Valentine
Gregory J Hannon
Affiliations
Soon will be listed here.
Abstract

RNA interference (RNAi) enables sequence-specific, experimentally induced silencing of almost any gene by tapping into innate regulatory mechanisms that are conserved among virtually all eukaryotes. In a typical RNAi experiment, an artificial silencing trigger directs the RNAi pathway toward a target that it would not normally recognize. This is most often an endogenous protein-coding gene, although some noncoding RNAs can also be silenced effectively. The artificial silencing trigger varies; this protocol uses synthetic small interfering RNAs (siRNAs). Lipofectamine 2000 is used to deliver the siRNAs into HEK293 cells. This lipid reagent has proven to be effective for many different cultured mammalian cell lines.

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