Date of Award

2018

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Area of Concentration

Biochemistry

Abstract

Many aspects have not yet been completely understood about rha-1 in C. elegans. In this thesis study, we attempted to build a CRISPR-Cas9 targeting vector to fuse GFP to the C-terminus of the rha-1 gene with the aim to acquire broader knowledge of the expression and functions of rha-1 in C. elegans. The CRISPR-Cas9 targeting vectors tagged by GFP at the C-terminus of the rha-1 gene were constructed using multiple DNA assembly tools such as: SapTrap Assembly, Gibson Assembly and Hi-Fi Assembly. Together, the obtained findings confirmed that the attempts to generate these CRISPR-Cas9 targeting vectors were unsuccessful due to numerous possible reasons. As a result, future work should primarily focus on developing an alternative way to tackle the inefficiency of constructing CRISPR-Cas targeting vectors for the rha-1 gene.

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