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Screening for gene function in chicken embryo using RNAi and electroporation

Nature Biotechnology volume 21, pages 93–96 (2003)Cite this article

An Erratum to this article was published on 01 February 2003

Abstract

In the postgenomic era the elucidation of the physiological function of genes has become the rate-limiting step in the quest to understand the development and function of living organisms. Gene functions cannot be determined by high-throughput methods but require analysis in the context of the entire organism. This is particularly true in the developing vertebrate nervous system1. Because of its easy accessibility in the egg, the chicken embryo has been the model of choice for developmental in vivo studies. However, its usefulness has been hampered by a lack of methods for genetic manipulation. Here we describe an approach that could compensate for this disadvantage. By combining gene silencing by dsRNA2 (through RNA interference, RNAi) with in ovo electroporation3,4, we developed an efficient method to induce loss of gene function in vivo during the development of the chicken CNS. This method opens new possibilities for studying gene function not only by gain-of-function but also by loss-of-function approaches and therefore represents a new tool for functional genomics.

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Figure 1: Demonstration of the efficiency of RNAi for interference with gene function in vivo.
Figure 2: In ovo RNAi produces specific and reproducible phenotypes in the embryonic chicken spinal cord.
Figure 3: A cDNA fragment is sufficient for the analysis of gene function in vivo by RNAi.

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References

  1. Lo, D.C. Challenges for neuroscience in a post-genome world. Nat. Neurosci. 4, 1153–1154 (2001).

    Article  CAS  Google Scholar 

  2. Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811 (1998).

    Article  CAS  Google Scholar 

  3. Muramatsu, T., Mizutani, Y., Ohmori, Y. & Okumura, J. Comparison of three nonviral transfection methods for foreign gene expression in early chicken embryos in ovo. Biochem. Biophys. Res. Commun. 230, 376–380 (1997).

    Article  CAS  Google Scholar 

  4. Itasaki, N., Bel-Vialar, S. & Krumlauf, R. 'Shocking' developments in chick embryology: electroporation and in ovo gene expression. Nat. Cell Biol. 1, E203–E207 (1999).

    Article  CAS  Google Scholar 

  5. Müller, U. Ten years of gene targeting: targeted mouse mutants, from vector design to phenotype analysis. Mech. Dev. 82, 3–21 (1999).

    Article  Google Scholar 

  6. Tang, J., Rutishauser, U. & Landmesser, L. Polysialic acid regulates growth cone behavior during sorting of motor axons in the plexus region. Neuron 13, 405–414 (1994).

    Article  CAS  Google Scholar 

  7. Stoeckli, E.T. & Landmesser, L.T. Axonin-1, NrCAM, and NgCAM play different roles in the in vivo guidance of chick commissural neurons. Neuron 14, 1165–1179 (1995).

    Article  CAS  Google Scholar 

  8. Burstyn-Cohen, T. et al. F-spondin is required for accurate pathfinding of commissural axons at the floor plate. Neuron 23, 233–246 (1999).

    Article  CAS  Google Scholar 

  9. Perrin, F.E., Rathjen, F.G. & Stoeckli, E.T. Distinct subpopulations of sensory afferents require F11 or axonin-1 for growth to their target layers within the spinal cord of the chick. Neuron 30, 707–723 (2001).

    Article  CAS  Google Scholar 

  10. Momose, T. et al. Efficient targeting of gene expression in chick embryos by microelectroporation. Dev. Growth Differ. 41, 335–344 (1999).

    Article  CAS  Google Scholar 

  11. Swartz, M., Eberhart, J., Mastick, G.S. & Krull, C.E. Sparking new frontiers: using in vivo electroporation for genetic manipulations. Dev. Biol. 233, 13–21 (2001).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  13. 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).

    Article  CAS  Google Scholar 

  14. Zhao, Z., Cao, Y., Li, M. & Meng, A. Double-stranded RNA injection produces nonspecific defects in zebrafish. Dev. Biol. 229, 215–223 (2001).

    Article  CAS  Google Scholar 

  15. Stoeckli, E.T. & Landmesser, L.T. Axon guidance at choice points. Curr. Opin. Neurobiol. 8, 73–79 (1998).

    Article  CAS  Google Scholar 

  16. Ikonomov, O.C., Kulesa, M.C., Shisheva, A.C. & Jacob, M.H. Innervation and target tissue interactions induce Rab-GDP dissociation inhibitor (GDI) expression during peripheral synapse formation in developing chick ciliary ganglion neurons in situ. J. Neurosci. 18, 6331–6339 (1998).

    Article  CAS  Google Scholar 

  17. Stenmark, H. & Olkkonen, V.M. The Rab GTPase family. Genome Biol. 2, Reviews 3007.1–3007.7 (2001).

    Article  Google Scholar 

  18. Geyer, M. & Wittinghofer, A. GEFs, GAPs, GDIs and effectors: taking a closer (3D) look at the regulation of Ras-related GTP-binding proteins. Curr. Opin. Struct. Biol. 7, 786–792 (1997).

    Article  CAS  Google Scholar 

  19. Perrin, F.E. & Stoeckli E.T. Use of lipophilic dyes in studies of axonal pathfinding in vivo. Microsc. Res. Tech. 48, 25–31 (2000).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Monika Mielich for excellent technical assistance. This work was carried out at the University of Basel, Institute of Zoology, and was supported by the Swiss National Science Foundation, the Ott Foundation, and the Human Frontier Science Program Organization.

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  1. University of Zurich, Institute of Zoology, Winterthurerstrasse 190, Zurich, CH-8057, Switzerland

    Vladimir Pekarik, Dimitris Bourikas, Nicola Miglino, Pascal Joset, Stephan Preiswerk & Esther T. Stoeckli

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  1. Vladimir Pekarik
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Correspondence to Esther T. Stoeckli.

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The authors declare no competing financial interests.

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Pekarik, V., Bourikas, D., Miglino, N. et al. Screening for gene function in chicken embryo using RNAi and electroporation. Nat Biotechnol 21, 93–96 (2003). https://doi.org/10.1038/nbt770

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