Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization

William K F Tse*, Yun Jin Jiang

*Corresponding author for this work

Research output: Chapter in book/report/conference proceedingChapterpeer-review

11 Citations (Scopus)


In order to unfold the function of genes, solely performing mRNA over-expression is not enough nowadays. Traditional protein expression experiments, such as Western blotting and immunohistochemical staining, could only provide researchers the changes of expression levels and/or location of their targets. To make a more strong and convincing statement about gene function, it is necessary to perform both "gain-of-function" and "loss-of-function" studies. Both assays can be performed easily by transfecting DNA plasmid and siRNA in cell culture system; while in zebrafish, mRNA and morpholino (MO) microinjection can serve similar purposes. It is common for the zebrafish community to carry out microinjection experiments to explore a gene function. Instead of making a single knockdown/over-expression of a gene, we foresee that more and more large-scale screens on certain protein families will be performed in the future. Here, based on our previous experience in zebrafish "loss-of-function" screening on deubiquitylating enzymes, we describe a general work flow, from morpholino designation, in situ hybridization, to data analysis, as a reference for researchers who may be interested in a similar screen.

Original languageEnglish
Title of host publicationFunctional Genomics
Subtitle of host publicationMethods and Protocols
EditorsMichael Kaufmann, Claudia Klinger
PublisherSpringer New York
Number of pages11
ISBN (Electronic)9781617794247
ISBN (Print)9781617794230, 9781493962280
Publication statusPublished - 4 Nov 2011

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Scopus Subject Areas

  • Molecular Biology
  • Genetics

User-Defined Keywords

  • "Loss-of-function" screening
  • Deubiquitylating enzymes
  • In situ hybridization
  • Morpholino
  • Zebrafish


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