Analysis of a novel regulatory mechanism of DNA damage repair using the combination of proteomics and genetics approarch

2015 Fiscal Year Final Research Report

• Project/Area Number: 26740018
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation and chemicals
• Research Institution: Kyoto University
• Principal Investigator: Sasanuma Hiroyuki 京都大学, 医学(系)研究科(研究院), 准教授 (00531691)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: DNA損傷 / プロテオミクス / ゲノム編集 / 相同組換え / DNA二重鎖切断修復

Outline of Final Research Achievements

During the period, I have already established the protocol of mass-spectrometry analysis using DT40 and human Tk6 B cell lines. Initially, I introduced 3flag tag at N-terminal region at first exon of BRCA1 using CRISPR/Cas9 genome editing. After pulling down 3FLAG-BRCA1 from whole cell extract, I analyzed the interactors of BRCA1, which plays a important role in DNA damage repair. I successfully identified many facotrs involved in transcription and chromosome segregation. Additionally, I identified the several novel proteins interacting with BRCA1. To investigate the function of novel BRCA1-interactors in DNA metabolism (DNA repair, replication, recombination), I continue generating the mutants of BRCA1 interactors identified here using human B cell line by genome editing technology. It is thought that the aim of this project was achieved.

Free Research Field

DNA損傷修復