|ZFIN ID: ZDB-PUB-170912-6|
Shifting Zebrafish Lethal Skeletal Mutant Penetrance by Progeny Testing
Brooks, E.P., Nichols, J.T.
|Source:||Journal of visualized experiments : JoVE (127): (Journal)|
|Registered Authors:||Brooks, Elliott, Nichols, James Tucker|
|PubMed:||28892034 Full text @ J. Vis. Exp.|
Brooks, E.P., Nichols, J.T. (2017) Shifting Zebrafish Lethal Skeletal Mutant Penetrance by Progeny Testing. Journal of visualized experiments : JoVE. (127).
ABSTRACTZebrafish mutant phenotypes are often incompletely penetrant, only manifesting in some mutants. Interesting phenotypes that inconsistently appear can be difficult to study, and can lead to confounding results. The protocol described here is a straightforward breeding paradigm to increase and decrease penetrance in lethal zebrafish skeletal mutants. Because lethal mutants cannot be selectively bred directly, the classic selective breeding strategy of progeny testing is employed. This method also includes protocols for Kompetitive Allele Specific PCR (KASP) genotyping zebrafish and staining larval zebrafish cartilage and bone. Applying the husbandry strategy described here can increase the penetrance of an interesting skeletal phenotype enabling more reproducible results in downstream applications. In addition, decreasing the mutant penetrance through this selective breeding strategy can reveal the developmental processes that most crucially require the function of the mutated gene. While the skeleton is specifically considered here, we propose that this methodology will be useful for all zebrafish mutant lines.
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