Evaluating the role of cis-regulatory tandem DNA repeats in human disease and evolution

2023 Fiscal Year Final Research Report

• Project/Area Number: 21H02460
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Kyoto University
• Principal Investigator: Woltjen Knut 京都大学, iPS細胞研究所, 准教授 (50589489)
• Co-Investigator(Kenkyū-buntansha): 川路 英哉 公益財団法人東京都医学総合研究所, ゲノム医学研究センター, 副センター長 (20525406) ; 依馬 正次 滋賀医科大学, 動物生命科学研究センター, 教授 (60359578) ; 井上 詞貴 京都大学, 高等研究院, 特定准教授 (60525369)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ゲノム編集 / ゲノム解析 / DNAリピート / ヒトiPS細胞

Outline of Final Research Achievements

Repetitive sequences are poorly understood regions of the human genome. Variable Number Tandem Repeats (VNTRs) are strings of DNA repeats that change in number between individuals. These changes can affect gene expression and may cause disease. To understand these relationships, we developed a novel gene editing technique to change VNTR copy numbers in human cells. After characterizing the copy number differences of 5 naturally occurring VNTRs across 22 human iPS cell lines, we used CRISPR-Cas9 to cut each repeat, triggering a cellular DNA repair that reduces repeat number to one. We repeated this process but protecting some repeats from being cut, enabling the generation of cells with intermediate repeat numbers, something which has never been previously achieved. We used this method to generate iPS cells with VNTRs of various copy numbers to model human disease. With these novel tools, we have begun to study VNTRs more broadly across the human genome and in non-human primate models.

Free Research Field

ゲノム生物学

Academic Significance and Societal Importance of the Research Achievements

VNTR copy-number editing is a new gene editing technique that allows scientists to study causal relationships between repetitive DNA and human disease. Achieving this in human iPS cells enables disease modeling of a new variety of genetic disorders in virtually all cell types of the body