Authors:

Morgan Stephens

Date Created:

Not specified

Course Title:

Professor:

Not specified

About Paper:

Would closure is driven by the actomyosin ring, through the recruitment of actin filaments and myosin motor proteins to the damaged site. Myosin itself is known to provide the contractive force behind this process. In our zebrafish model, we aim to observe this process in by targeting the non- muscle myosin gene myh9a that is involved in tail contraction, as the mechanisms that are utilized by myh9a in the process of wound closure are not understood. This research aims to better understand these functions of myh9a. This study utilizes a two-step Knock In (KI) method with the PhiC31-CRISPR Cas9 recombination system in examining myh9a receptor interactions. The first step is the generation of an attp KI line, followed by the injection of an attB plasmid into the attp line. This integration is directed through site-specific attp-attB DNA recombination and is performed by inducing DSBs to the last few introns to lower disruption in the translation of the endogenous protein. Endogenous gene tagging can be used to examine these expressions and to study protein localization within a cell. Through utilizing myh9a that has been tagged with a green fluorescent protein (GFP) or red fluorescent protein (RFP), these interactions of myh9a with receptors and proteins can be tracked in the epithelial skin layers. Overall, the aim of this work is to observe and determine the roles of myosin gene myh9a in Zebrafish wound closure. Keywords: CRISPR-Cas9; Knock-In Lines; Myh9a; Wound Closure

Source:

Purdue University / 2025

Topics:

No topics listed

Co-authors:

Morgan Stephens