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Every cell has to undergo the cell cycle process before it can divide or differentiate. The cell cycle is an error-prone complex process where mutations could happen. This study aimed to characterize the nervous system and blood cells of the speed bump mutant in zebrafish. This study also aimed to do gene characterization of the wee1 CRISPR mutant. The speed bump mutant is due to a mutation in the wee1 gene. Lack of the wee1 gene will cause no Wee1 Kinase, which inhibits the MPF, causing cells to go into mitosis too early. To fulfill these objectives, this study used in situ hybridization with neural probes and blood probes, antibody staining, as well as sanger sequencing. It is found that, like other cell cycle mutants, neural cells are differentiated in spbti279 mutants, but, unlike other cell cycle mutants, spbti279 mutants are still able to make neuroblasts (stem cells that have decided to become neurons) even at the later stages of development. It is also found that these spbti279 mutants do not produce red blood cells. This study also found that the wee1 CRISPR mutants all have more severe phenotypes compared to the spbti279 mutant and that frameshift mutations cause the wee1 CRISPR mutants to lose the Wee1 protein kinase domain. In spbti279 mutants, the Wee1 protein kinase domain still exists though it lost the last ten amino acids. These results indicate that the spbti279 mutants are less likely to have a loss of function mutation and more likely to have hypomorph mutation. The wee1 CRISPR mutants are likely to have a complete loss of function. The information gathered in this study will allow a better understanding about the wee1 gene as well as the Wee1 Kinase domain during development.
Karyadi, Nathania, "Is the Zebrafish speed bump Mutant a Mutation in wee1? And Is It a Complete Loss of Function?" (2022). Honors Theses. 3552.
Honors Thesis-Open Access