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Manipulation of Gene Expression During Zebrafish Embryonic Development Using Transient Approaches

  • Protocol
Wnt Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 469))

Abstract

The rapid embryonic development and high fecundity of zebrafish contribute to the great advantages of this model for the study of developmental genetics. Transient disruption of the normal function of a gene during development can be achieved by microinjecting mRNA, DNA or short chemically stabilized anti-sense oligomers, called morpholinos (MOs), into early zebrafish embryos. The ensuing develop ment of the microinjected embryos is observed over the following hours and days to analyze the impact of the microinjected products on embryogenesis. Compared to stable reverse genetic approaches (sta ble transgenesis, targeted mutants recovered by TILLING), these transient reverse genetic approaches are vastly quicker, relatively affordable, and require little animal facility space. Common applications of these methodologies allow analysis of gain-of-function (gene overexpression or dominant active), loss-of-function (gene knock down or dominant negative), mosaic analysis, lineage-restricted studies and cell tracing experiments. The use of these transient approaches for the manipulation of gene expression has improved our understanding of many key developmental pathways including both the Wnt/β -catenin and Wnt/PCP pathways, as covered in some detail in Chapter 17 of this book. This chapter describes the most common and versatile approaches: gain of function and loss of function using DNA and mRNA injections and loss of function using MOs.

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Acknowledgments

The authors thank Pierre Smith, Department of Surgery, Royal Melbourne Hospital for photography and Andrew Badrock from the Heath laboratory for appearing in Fig. 19.1 and setting up the materials for the other figures.

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Authors and Affiliations

  1. Hubrecht Institute for Developmental Biology and Stem Cell Research, Uppsalalaan 8, 3584, CT Utrecht, The Netherlands

    Benjamin M. Hogan

  2. School of Biological Sciences, Monash University, Clayton, Victoria, Australia

    Heather Verkade

  3. The Walter and Eliza Hall Institute of Medical Research, Parkville, Melbourne, Victoria, Australia

    Graham J. Lieschke

  4. Ludwig Institute of Cancer research, Parkville, Victoria, Australia

    Joan K. Heath

Authors
  1. Benjamin M. Hogan
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  2. Heather Verkade
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  3. Graham J. Lieschke
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  4. Joan K. Heath
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Editor information

Editors and Affiliations

  1. University of Melbourne, VIC 3010, Parkville, Victoria, Australia

    Elizabeth Vincan

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Hogan, B.M., Verkade, H., Lieschke, G.J., Heath, J.K. (2008). Manipulation of Gene Expression During Zebrafish Embryonic Development Using Transient Approaches. In: Vincan, E. (eds) Wnt Signaling. Methods in Molecular Biology, vol 469. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-469-2_19

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  • DOI: https://doi.org/10.1007/978-1-60327-469-2_19

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60327-468-5

  • Online ISBN: 978-1-60327-469-2

  • eBook Packages: Springer Protocols

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