Date of Award

8-1-2017

Degree Name

Master of Science

Department

Molecular Biology Microbiology and Biochemistry

First Advisor

Davie, Judith

Abstract

Myogenesis, the formation of muscular tissues, requires the precise expression of myogenic regulatory factors (MRFs), whose expression regulates transcription of muscle specific genes. TBX3 is a member of the T-box family of transcription factors that is expressed throughout myogenesis and is up regulated. Our aim was to understand the role of TBX3 in myogenesis. Therefore, we knocked out TBX3 from C2C12 cell lines using the Cripr/Cas9 system which is an RNA-mediated system that has been proven to be an efficient way to perform a targeted and precise genome editing in living cells. Results form qRT-PCR show that there is a decrease in the expression level of the muscle-specific genes (TNNI2, Mylpf, and Lmod2). We also tested the effect of knocking out TBX3 on the expression of the myogenic regulatory factor (myogenin), and we found out that myogenin does not become up regulated upon differentiation in knockout cells. These results suggest that Knocking out TBX3 results in delayed differentiation of skeletal muscle cells and is required for normal progression of myogenesis.

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