Editors' ChoiceMethodology

A Knock-Out Method with Double-Stranded RNA

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Science's STKE  08 Feb 2000:
Vol. 2000, Issue 18, pp. tw1
DOI: 10.1126/stke.2000.18.tw1

Double-stranded RNA injection has been used successfully in the nematode to specifically disrupt expression from target genes. The mechanism of this type of inhibition is not understood; however, repression of transcription of the targeted gene persists through many cell cycles in Caenorhabditis elegans. The double-stranded RNA can be injected at the single cell stage allowing efficient inhibition of transcription of a specific gene in all cells in the developing embryo. Li et al. demonstrate that injection of double-stranded RNA effectively inhibits transcription of target genes in zebrafish embryos. Expression from a microinjected plasmid was suppressed by injection of double-stranded RNA. Additionally, expression of endogenous proteins was suppressed by injection of targeted double-stranded RNA. This procedure reproduced null phenotypes associated with loss of the targeted gene or genes. Double-stranded RNA-mediated gene silencing in zebrafish provides another tool in a genetically tractable system for studying signal transduction pathways.

Li, Y.-X., Farrell, M.J., Liu, R., Mohanty, N., and Kirby, M.L. (2000) Double-stranded RNA injection produces null phenotypes in zebrafish. Dev. Biol. 217: 394-405. [Abstract]

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