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

Ka Fai William Tse, Yun Jin Jiang

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Citations (Scopus)

Abstract

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
Pages321-331
Number of pages11
DOIs
Publication statusPublished - Jan 2 2012
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume815
ISSN (Print)1064-3745

Fingerprint

Morpholinos
Zebrafish
In Situ Hybridization
Microinjections
Enzymes
Research Personnel
Genes
Messenger RNA
Workflow
Small Interfering RNA
Proteins
Plasmids
Cell Culture Techniques
Western Blotting
Staining and Labeling
Gene Expression
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

Tse, K. F. W., & Jiang, Y. J. (2012). Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization. In M. Kaufmann, & C. Klinger (Eds.), Functional Genomics: Methods and Protocols (pp. 321-331). (Methods in Molecular Biology; Vol. 815). https://doi.org/10.1007/978-1-61779-424-7_24

Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization. / Tse, Ka Fai William; Jiang, Yun Jin.

Functional Genomics: Methods and Protocols. ed. / Michael Kaufmann; Claudia Klinger. 2012. p. 321-331 (Methods in Molecular Biology; Vol. 815).

Research output: Chapter in Book/Report/Conference proceedingChapter

Tse, KFW & Jiang, YJ 2012, Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization. in M Kaufmann & C Klinger (eds), Functional Genomics: Methods and Protocols. Methods in Molecular Biology, vol. 815, pp. 321-331. https://doi.org/10.1007/978-1-61779-424-7_24
Tse KFW, Jiang YJ. Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization. In Kaufmann M, Klinger C, editors, Functional Genomics: Methods and Protocols. 2012. p. 321-331. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-424-7_24
Tse, Ka Fai William ; Jiang, Yun Jin. / Functional screen of zebrafish deubiquitylating enzymes by morpholino knockdown and in situ hybridization. Functional Genomics: Methods and Protocols. editor / Michael Kaufmann ; Claudia Klinger. 2012. pp. 321-331 (Methods in Molecular Biology).
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