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.

Accessibility Status

Searchable text

Creative Commons License

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

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.