Stable suppression of gene expression by RNAi in mammalian cells

Paddison, P. J., Caudy, A. A., Hannon, G. J. (2002) Stable suppression of gene expression by RNAi in mammalian cells. Proceedings of the National Academy of Sciences of the United States of America, 99 (3). pp. 1443-1448. ISSN 00278424 (ISSN)

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Abstract

In a diverse group of organisms including plants, Caenorhabditis elegans, Drosophila, and trypanosomes, double-stranded RNA (dsRNA) is a potent trigger of gene silencing. In several model systems, this natural response has been developed into a powerful tool for the investigation of gene function. Use of RNA interference (RNAi) as a genetic tool has recently been extended to mammalian cells, being inducible by treatment with small, ≈22-nt RNAs that mimic those produced in the first step of the silencing process. Here, we show that some cultured murine cells specifically silence gene expression upon treatment with long dsRNAs (≈500 nt). This response shows hallmarks of conventional RNAi including silencing at the posttranscriptional level and the endogenous production of ≈22-nt small RNAs. Furthermore, enforced expression of long, hairpin dsRNAs induced stable gene silencing. The ability to create stable "knock-down" cell lines expands the utility of RNAi in mammalian cells by enabling examination of phenotypes that develop over long time periods and lays the groundwork for by using RNAi in phenotype-based, forward genetic selections.

Item Type:	Paper
Uncontrolled Keywords:	double stranded RNA animal cell article controlled study gene expression regulation gene function gene silencing genetic selection knockout gene mammal cell nonhuman phenotype priority journal RNA transcription Animals Cells, Cultured Gene Expression Regulation, Neoplastic Genes, Reporter Luciferases Mammals Mice RNA, Antisense RNA, Double-Stranded RNA, Messenger RNA, Small Interfering Stem Cells Transfection Tumor Cells, Cultured Animalia Caenorhabditis Caenorhabditis elegans Mammalia Murinae
Subjects:	bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification Investigative techniques and equipment > RNAI bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi organs, tissues, organelles, cell types and functions > cell types and functions bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function organism description > animal > mammal
CSHL Authors:	Caudy, Amy A. Hannon, Gregory J. Paddison, Patrick J.
Communities:	CSHL labs > Hannon lab School of Biological Sciences > Publications
Depositing User:	Matt Covey
Date:	2002
Date Deposited:	09 Jan 2013 16:57
Last Modified:	10 Sep 2019 19:06
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/26465

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