Submission Type
Poster
Start Date
April 2021
Abstract
In Zebrafish, the chromosome assembly factor 1b (chaf1b) gene is in part responsible for the development of the eye. In homozygous chaf1bt24412 mutants retinal cell death is promoted through cell-death promoting activity of the gene, tumor suppressor protein p53 (tp53), resulting in a small-eye phenotype. Another allele chaf1bnt2, was found to also result in the small-eye phenotype when in a homozygous state. We found that knockdown of Tp53 protein via morpholino antisense oligonucleotide injection of 1-2 cell stage embryos failed to rescue retinal cell death of chaf1bnt2 homozygous mutants as detected by TUNEL labeling. Because morpholinos may fail to fully inhibit target gene function we crossed carrier fish heterozygous for both the nt2 and a cell-death induction deficient allele of tp53 (zdf1) and compared double homozygous mutants to siblings which had chaf1b mutant homozygosity and functional tp53. Restriction fragment length polymorphism analysis was used to verify zygosity of the nt2 and zdf1 alleles. We found that loss of tp53 function failed to rescue the chaf1bnt2 small-eye phenotype.
Recommended Citation
Parks, Alex, "216— Loss of Function Mutation for tp53 Does Not Rescue the chaf1bnt2 Small-eye Phenotype in Danio rerio" (2021). GREAT Day Posters. 104.
https://knightscholar.geneseo.edu/great-day-symposium/great-day-2021/posters-2021/104
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216— Loss of Function Mutation for tp53 Does Not Rescue the chaf1bnt2 Small-eye Phenotype in Danio rerio
In Zebrafish, the chromosome assembly factor 1b (chaf1b) gene is in part responsible for the development of the eye. In homozygous chaf1bt24412 mutants retinal cell death is promoted through cell-death promoting activity of the gene, tumor suppressor protein p53 (tp53), resulting in a small-eye phenotype. Another allele chaf1bnt2, was found to also result in the small-eye phenotype when in a homozygous state. We found that knockdown of Tp53 protein via morpholino antisense oligonucleotide injection of 1-2 cell stage embryos failed to rescue retinal cell death of chaf1bnt2 homozygous mutants as detected by TUNEL labeling. Because morpholinos may fail to fully inhibit target gene function we crossed carrier fish heterozygous for both the nt2 and a cell-death induction deficient allele of tp53 (zdf1) and compared double homozygous mutants to siblings which had chaf1b mutant homozygosity and functional tp53. Restriction fragment length polymorphism analysis was used to verify zygosity of the nt2 and zdf1 alleles. We found that loss of tp53 function failed to rescue the chaf1bnt2 small-eye phenotype.
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