| Project/Area Number |
23KF0041
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| Research Category |
Grant-in-Aid for JSPS Fellows
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| Allocation Type | Multi-year Fund |
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| Section | 外国 |
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| Review Section |
Basic Section 43050:Genome biology-related
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| Research Institution | Kyoto University |
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Principal Investigator |
ウォルツェン クヌート 京都大学, iPS細胞研究所, 准教授 (50589489)
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| Co-Investigator(Kenkyū-buntansha) |
MARTINEZ GALVEZ GABRIEL 京都大学, iPS細胞研究所, 外国人特別研究員
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| Project Period (FY) |
2023-04-25 – 2024-03-31
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| Project Status |
Completed (Fiscal Year 2023)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2023: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | DNAリピート / ゲノム編集 / CRISPR-Cas9 / iPS細胞 / レポーターアッセイ |
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| Outline of Research at the Start |
Tandem repeat copy-number variations have been correlated with changes in gene expression and are probable causes of human disease. We are developing gene editing and computational biology methods to efficiently generate repeat-variant disease models in human induced pluripotent stem (iPS) cells. Moreover, we are developing technologies to produce intermediate copy-number variants. As proof-of-concept, we completed the generation of a repeat-copy variants in iPS cells at two loci with known clinically relevance, and aim to apply these methods genome-wide.
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| Outline of Annual Research Achievements |
Variable Number Tandem Repeats (VNTRs) are repetitive DNA sequences that differ in number between individuals. VNTR copy number is shown to correlate with changes in gene expression, and are probable causes of human disease. To understand these relationships, we developed a new bioinformatic pipeline to identify, analyze, and design gene editing strategies for VNTRs, which has never been previously achieved. We characterized 5 VNTRs for copy-number polymorphisms across 22 human iPS cell lines. We designed CRISPR-Cas9 strategies that target and cut each repeats, triggering cellular DNA repair that reduces the repeat number to one. Moreover, we developed a novel gene editing strategy to control VNTR copy-number to generate iPS cells with VNTRs of various copy numbers for disease modeling.
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