PUBLICATION
Undertaking a successful gynogenetic haploid screen in zebrafish
- Authors
- Layton, J.E.
- ID
- ZDB-PUB-090422-23
- Date
- 2009
- Source
- Methods in molecular biology (Clifton, N.J.) 546: 31-44 (Chapter)
- Registered Authors
- Layton, Judy E.
- Keywords
- Mutagenesis, ENU, Gynogenetic, Haploid, Screen Zebrafish
- MeSH Terms
-
- DNA/drug effects
- DNA/radiation effects
- Genetic Testing/methods*
- Animals
- Ultraviolet Rays
- Mutagenesis
- Heterozygote
- Fertilization in Vitro
- Female
- Alkylating Agents/administration & dosage
- Haploidy*
- Zebrafish/embryology
- Zebrafish/genetics*
- Ethylnitrosourea/administration & dosage
- Genetic Engineering/methods
- Male
- Spermatozoa/drug effects
- Spermatozoa/radiation effects
- PubMed
- 19378096 Full text @ Meth. Mol. Biol.
Citation
Layton, J.E. (2009) Undertaking a successful gynogenetic haploid screen in zebrafish. Methods in molecular biology (Clifton, N.J.). 546:31-44.
Abstract
Chemical mutagenesis using N-ethyl-N-nitrosourea is the current method of choice for dense mutagenesis in zebrafish. Methods are available for both pre-meiotic and post-meiotic sperm mutagenesis; in this chapter, pre-meiotic mutagenesis is described. Mutated males are crossed with untreated females to create an F1 generation that is heterozygous for the mutations. The F1 females can be screened directly by making haploid embryos using in vitro fertilization (IVF) with ultraviolet (UV)-irradiated sperm. This approach requires substantially fewer fish and less aquarium space than the classical F2 generation screen and is feasible for a small research group. Production of haploid embryos is described in detail.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping