Honors Theses

Date of Award

Spring 5-9-2020

Document Type

Undergraduate Thesis

Department

Biology

First Advisor

Joshua Bloomekatz

Second Advisor

Sarah Liljegren

Third Advisor

Gregg Roman

Relational Format

Dissertation/Thesis

Abstract

Zebrafish are a good model organism in which to identify genes important for heart development because most human genes have a zebrafish ortholog and genes in zebrafish are relatively simple to modify. ccdc141 is a gene with unknown function, which we originally found to be highly expressed in cardiac cells in mouse embryos. In this project, we are seeking to determine the function of ccdc141 using zebrafish. To perform this experiment, we are targeting ccdc141 for mutation using CRISPR/Cas9. In this project we have analyzed ccdc141 to determine regions of conservation, designed CRISPR guides to target those regions (exon 2 or 3), injected ribonucleoprotein (RNP) complexes containing the generated gRNA and Cas9 into zebrafish embryos, and designed and executed assays to assess the efficacy of those injections. Once a DNA cut is made using gRNA/Cas9, the cell repairs those cuts during which a mutation may be introduced. The mutation introduced may disrupt regular gene function, allowing for a chance to view how the organism functions without a functional ccdc141 gene. The expectation is that ccdc141 is necessary for heart function, including the early stages of heart tube formation, namely cardiac fusion and cardiac cell movement. Although we were unable to determine whether a phenotype was created, we did determine that we successfully created a double-strand break in ccdc141.

Creative Commons License

Creative Commons Attribution-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-No Derivative Works 4.0 License.

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