Date Approved

2023

Degree Type

Open Access Senior Honors Thesis

Department or School

Biology

First Advisor

Robert Winning, Ph.D.

Second Advisor

Aaron Liepman, Ph.D.

Third Advisor

Natalie Dove, Ph.D.

Abstract

Cellular communication is important in embryonic development. It facilitates timely migration and maturation of cells to produce a fully functional organism using cell signaling pathways. These pathways involve receptors, ligands, and intracellular enzymes to produce different effects in the cell. One such pathway, the EphA4 signaling pathway, is involved in altering the cytoskeleton of eukaryotic cells. LIM kinases (LIMKs) are enzymes that regulate important components of the cytoskeleton. We hypothesized involvement of LIMKs in the EphA4 pathway and investigated their involvement in X. laevis embryonic development as a first step toward evaluating this hypothesis. We mutated LIMK genes in frog embryos using CRISPR/Cas9, and used microscopy to examine embryo development, to look for an association between mutation and altered embryonic morphology. We show that while CRISPR/Cas9 is a valid strategy to introduce mutations in LIMK genes of frog embryos, obvious defects relating to embryonic development were not observed. We discuss explanations and potential pitfalls of this approach and suggest alternatives for future studies.

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