Forward genetic screen using zebrafish to identify new genes involved in myelination

Linde Kegel, Maria Rubio, Rafael G. Almeida, Silvia Benito, Anna Klingseisen, David A. Lyons*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Zebrafish are now well established as the preeminent vertebrate model with which to carry out gene discovery/forward genetic screens to identify the molecular genetic basis of biological processes. Gene discovery screens in zebrafish have already provided novel insight into mechanisms of glial cell development and function. The vast majority of genetic screens in zebrafish are based around a three generation screen that starts with the random induction of mutations in adult males using the chemical mutagen ENU. Here we outline the methods that underlie this type of screen, detailing each step, from ENU mutagenesis, through the breeding schemes required to recover homozygous mutant animals in subsequent generations, the screening procedure itself, with a focus on the analysis of myelinating glia, and the subsequent confirmation of mutant phenotypes.

Original languageEnglish
Title of host publicationOligodendrocytes
Subtitle of host publicationMethods and Protocols
EditorsDavid A. Lyons, Linde Kegel
Place of PublicationNew York
PublisherHumana Press
Pages185-209
Number of pages25
Edition1
ISBN (Electronic)9781493990726
ISBN (Print)9781493990702
DOIs
Publication statusPublished - 1 Mar 2019

Publication series

NameMethods in Molecular Biology
Volume1936
ISSN (Print)1064-3745

Keywords / Materials (for Non-textual outputs)

  • ENU
  • Forward genetics screen
  • Myelin
  • Oligodendrocyte
  • Whole genome sequencing
  • Zebrafish

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