PUBLICATION

Forward Genetic Screen Using Zebrafish to Identify New Genes Involved in Myelination

Authors
Kegel, L., Rubio, M., Almeida, R.G., Benito, S., Klingseisen, A., Lyons, D.A.
ID
ZDB-PUB-190302-4
Date
2019
Source
Methods in molecular biology (Clifton, N.J.)   1936: 185-209 (Chapter)
Registered Authors
Almeida, Rafael, Kegel, Linde, Lyons, David A., Rubio, Maria
Keywords
ENU, Forward genetics screen, Myelin, Oligodendrocyte, Whole genome sequencing, Zebrafish
MeSH Terms
  • Animals
  • Breeding
  • Ethylnitrosourea/adverse effects*
  • Female
  • Genetic Testing/methods*
  • Homozygote
  • Male
  • Models, Animal
  • Mutation
  • Myelin Sheath/genetics*
  • Phenotype
  • Zebrafish/genetics*
  • Zebrafish Proteins/genetics*
PubMed
30820900 Full text @ Meth. Mol. Biol.
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.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping