The Tumour Suppressor Tuberin and DNA Damage Response
Standing
Undergraduate
Type of Proposal
Oral Research Presentation
Challenges Theme
Open Challenge
Faculty Sponsor
Dr. Lisa Porter, Dr. Elizabeth Fidalgo Da Silva
Proposal
The cell cycle is regulated by checkpoints that delay mitotic progression. DNA damage can be caused by an array of processes, like radiation and chemotherapy agents, leading to cellular mutations and carcinogenesis. Tuberin (TSC2), a tumour suppressor protein, regulates the G2/M transition in the cell cycle and negatively regulates protein synthesis and cell growth. Previously, our lab has shown that Tuberin regulates mitotic onset through cellular localization of the G2/M Cyclin, Cyclin B1. My project focuses on the Tuberin/Cyclin B1 complex and its role in G2/M arrest and DNA damage repair. In this study, two Tuberin null NIH-3T3 (mouse) cell lines will be created through CRISPR-Cas9 editing to define the role of Tuberin during DNA repair. Etoposide, a topoisomerase II drug, will be used to induce DNA damage. Cells will then be analyzed by flow cytometry and western blotting to assess their cell cycle profile, apoptotic levels, and TSC2 protein expression.
Grand Challenges
Viable, Healthy and Safe Communities
The Tumour Suppressor Tuberin and DNA Damage Response
The cell cycle is regulated by checkpoints that delay mitotic progression. DNA damage can be caused by an array of processes, like radiation and chemotherapy agents, leading to cellular mutations and carcinogenesis. Tuberin (TSC2), a tumour suppressor protein, regulates the G2/M transition in the cell cycle and negatively regulates protein synthesis and cell growth. Previously, our lab has shown that Tuberin regulates mitotic onset through cellular localization of the G2/M Cyclin, Cyclin B1. My project focuses on the Tuberin/Cyclin B1 complex and its role in G2/M arrest and DNA damage repair. In this study, two Tuberin null NIH-3T3 (mouse) cell lines will be created through CRISPR-Cas9 editing to define the role of Tuberin during DNA repair. Etoposide, a topoisomerase II drug, will be used to induce DNA damage. Cells will then be analyzed by flow cytometry and western blotting to assess their cell cycle profile, apoptotic levels, and TSC2 protein expression.