Manipulation of gene expression during zebrafish embryonic development using transient approaches

Benjamin M. Hogan, Heather Verkade, Graham J. Lieschke, Joan K. Heath

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

34 Citations (Scopus)


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.

Original languageEnglish
Title of host publicationWnt Signaling
Subtitle of host publicationVolume 2 Pathway Models
EditorsElizabeth Vincan
Place of PublicationNew York USA
PublisherHumana Press
Number of pages28
ISBN (Print)9781603274685
Publication statusPublished - 2008

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • DNA
  • Gain-of-function phenotype
  • Loss-of-function phenotype
  • Microinjection
  • Morpholino
  • Mutant
  • RNA
  • Zebrafish embryos

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