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Specific interference with gene function by double-stranded RNA in early mouse development

Nature Cell Biology volume 2, pages 70–75 (2000)Cite this article

Abstract

The use of double-stranded (ds) RNA is a powerful way of interfering with gene expression in a range of organisms, but doubts have been raised about whether it could be successful in mammals. Here, we show that dsRNA is effective as a specific inhibitor of the function of three genes in the mouse, namely maternally expressed c-mos in the oocyte and zygotically expressed E-cadherin or a GFP transgene in the preimplantation embryo. The phenotypes observed are the same as those reported for null mutants of the endogenous genes. These findings offer the opportunity to study development and gene regulation in normal and diseased cells.

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Figure 1: MmGFP dsRNA specifically abrogates MmGFP expression in MmGFP transgenic embryos.
Figure 2: Interference with expression of injected synthetic MmGFP mRNA.
Figure 3: Injection of E-cadherin dsRNA to the zygote reduces E-cadherin expression and perturbs the development of the injected embryos.
Figure 4: Incidence of cavity formation after injection of E-cadherin dsRNA into the zygote.
Figure 5: Injection of c-mos dsRNA into immature oocytes inhibits c-mos expression and causes parthenogenetic activation.
Figure 6: Incidence of spontaneous activation and fragmentation after injection of c-mos dsRNA into the germinal-vesicle-stage oocyte.

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Acknowledgements

We thank D. Glover for suggestions and support throughout the course of this work; J. Gurdon for the initial idea of injecting mRNA into embryos; A. McLaren for her suggestion of the use of E-cadherin as a marker; L. Larue for E-cadherin plasmids and antibody; B. Colledge for the c-mos plasmid; P.-Y. Bourillot for subcloning the c-mos construct and for helpful discussions; R. Weber for supportive discussions and help in initiating microinjection experiments; M. Evans for his enthusiasm and support with the initiation of this work; and B. Tom for the help with the statistical analysis. This work was supported by a CRC Grant to M.Z.-G., D. Glover and M. Evans. M.Z.-G. is a Senior Research Fellow of the Lister Institute for Preventive Medicine, and a Stanley Elmore Fellow of Sydney Sussex College.

Correspondence and requests for materials should be addressed to M.Z.-G.

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  1. Wellcome/CRC Institute and Department of Genetics, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR , UK

    Florence Wianny & Magdalena Zernicka-Goetz

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  1. Florence Wianny
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Correspondence to Magdalena Zernicka-Goetz.

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Wianny, F., Zernicka-Goetz, M. Specific interference with gene function by double-stranded RNA in early mouse development. Nat Cell Biol 2, 70–75 (2000). https://doi.org/10.1038/35000016

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