| 研究実績の概要 |
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. 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, which triggers cellular DNA repair and reduces the repeat number to one. We verified the repeat sequence with long-read nanopore technology. Moreover, we developed a new gene editing strategy to control VNTR copy-number reduction, which has never been previously achieved. We used this method to generate iPS cells with VNTRs of various copy numbers for disease modeling. With these novel tools, we have begun to study VNTRs more broadly across the human genome and in non-human primate models.
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