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Using Single Molecule Live Cell Imaging to Investigate the Role of RNF169 in the DNA Damage Response
Aastha Bahl, 3rd-year
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Abstract:
Human ring finger protein 169 (RNF169) plays an important role in mediating the DNA damage response, specifically in the response to DNA double strand breaks (DSBs).
DSBs in DNA elicit RNF168 to recruit the DNA repair protein 53BP1 as well as RNF169 through the deposition of a ubiquitin modification on histone H2A. The literature shows that RNF169 limits the accumulation of 53BP1, thereby attenuating 53BP1 activity. The purpose of this research is to investigate the way that RNF169 balances the DNA damage response. Through designing and utilizing CRISPR plasmids for endogenous genome editing of RNF169 in U2OS cells, I have generated HALO tagged RNF169 cell lines for live-cell single-molecule imaging studies. Using this genome-edited cell line, I can analyze the recruitment of RNF169 to DNA damage sites and define the molecular mechanism by which it counteracts 53BP1 recruitment.