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Shifting Zebrafish Lethal Skeletal Mutant Penetrance by Progeny Testing
Published on: September 1, 2017
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Shifting Zebrafish Lethal Skeletal Mutant Penetrance by Progeny Testing
Elliott P Brooks1, James T Nichols2
1Department of Craniofacial Biology, University of Colorado Anschutz Medical Campus.
Journal of Visualized Experiments : Jove
|September 12, 2017
Summary
This study presents a breeding strategy to control the penetrance of lethal zebrafish skeletal mutations. This method enhances reproducibility and aids in understanding gene function during development.
Area of Science:
- Developmental biology
- Genetics
- Zebrafish models
Background:
- Incomplete penetrance of zebrafish mutant phenotypes complicates research and yields inconsistent results.
- Lethal mutations pose unique challenges for selective breeding due to the inability to directly select affected individuals.
Purpose of the Study:
- To describe a breeding paradigm for manipulating penetrance in lethal zebrafish skeletal mutants.
- To enable more reproducible studies of incompletely penetrant phenotypes.
- To reveal critical developmental roles of mutated genes.
Main Methods:
- Utilized progeny testing, a classic selective breeding strategy, for lethal mutants.
- Incorporated Kompetitive Allele Specific PCR (KASP) genotyping for zebrafish.
- Included protocols for staining larval zebrafish cartilage and bone.
Main Results:
- The described husbandry strategy successfully increases penetrance of skeletal phenotypes.
- Decreasing penetrance revealed crucial developmental processes dependent on mutated gene function.
- The methodology is applicable beyond skeletal mutants to all zebrafish lines.
Conclusions:
- This straightforward breeding paradigm enhances the study of incompletely penetrant zebrafish mutants.
- The strategy improves experimental reproducibility and aids in dissecting gene function.
- The proposed methodology offers broad utility across various zebrafish mutant lines.
