Mutation Generation on the BRCA2 using Recombineering

Presenter: Samyak Jain, Global Public Health Scholars

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For my practicum project, I completed an internship through the Summer Internship Program in the at NCI-Frederick (National Cancer Institute) in the Mouse Cancer Genetics Lab. I was working under Arun Mishra, a post-doc in the lab. The lab focuses on BRCA, a crucial gene dealing with DNA damage, and is a known tumor suppressor. Those with mutations in this gene have an increased chance of developing breast and ovarian cancer. Thus, the lab is focused on understanding how BRCA2 mutations can change the genotype of the mice. As a lab intern, I was tasked with creating a mutation via 2-step hit-and-fix recombineering. The procedure involved me manipulating the germline cells in mice and inserting a mutated sequence to then grow into mice. My daily day in the lab consisted of me utilizing various laboratory techniques such as creating PCRs, gel-electrophoresis, genotyping mice tails, extracting and digesting DNA, and culturing cells. I learned how researchers can manipulate genes and analyze them for a mutation and also how a lab environment is. I also learned the importance of communication and sharing information within the lab setting. The major takeaway from this experience has also been the patience required to be in a lab environment. From a public health standpoint, cancer is one of the leading causes of death worldwide, so researching a gene that is known to cause cancer can help researchers identify causes, diagnosis, prevention, and treatment. Results from this research can be used to create awareness campaigns, cancer screening programs, research of risk factors, and the development of treatments. This research can also show disparities such as how racial minorities and low-income communities are more likely to experience cancer and have worse cancer outcomes.