Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans

Abstract

Experimental introduction of RNA into cells can be used in certain biological systems to interfere with the function of an endogenous gene1,2. Such effects have been proposed to result from a simple antisense mechanism that depends on hybridization between the injected RNA and endogenous messenger RNA transcripts. RNA interference has been used in the nematode Caenorhabditis elegans to manipulate gene expression3,4. Here we investigate the requirements for structure and delivery of the interfering RNA. To our surprise, we found that double-stranded RNA was substantially more effective at producing interference than was either strand individually. After injection into adult animals, purified single strands had at most a modest effect, whereas double-stranded mixtures caused potent and specific interference. The effects of this interference were evident in both the injected animals and their progeny. Only a few molecules of injected double-stranded RNA were required per affected cell, arguing against stochiometric interference with endogenous mRNA and suggesting that there could be a catalytic or amplification component in the interference process.

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Figure 1: Genes used to study RNA-mediated genetic interference in C.elegans.
Figure 2: Analysis of RNA-interference effects in individual cells.
Figure 3: Effects of mex-3 RNA interference on levels of the endogenous mRNA.

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Acknowledgements

We thank A. Grishok, B. Harfe, M. Hsu, B. Kelly, J. Hsieh, M. Krause, M. Park, W. Sharrock, T. Shin, M. Soto and H. Tabara for discussion. This work was supported by the NIGMS (A.F.) and the NICHD (C.M.), and by fellowship and career awards from the NICHD (M.K.M.), NIGMS (S.K.), PEW charitable trust (C.M.), American Cancer Society (C.M.), and March of Dimes (C.M.).

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Correspondence to Andrew Fire.

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Fire, A., Xu, S., Montgomery, M. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811 (1998). https://doi.org/10.1038/35888

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