2020 Fiscal Year Final Research Report
Analysis of lineage-specific chromosomal conformation by in vitro enChIP
| Project/Area Number |
18K06176
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 43050:Genome biology-related
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| Research Institution | Hirosaki University |
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Principal Investigator |
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | ChIP / ChIP-Seq / locus-specific ChIP / CRISPR / enChIP / in vitro enChIP / in vitro enChIP-Seq |
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| Outline of Final Research Achievements |
We previously developed in vitro enChIP, which is a biotechnology to isolate a target genomic region from cells. In this study, we combined in vitro enChIP with next-generation sequencing analysis (in vitro enChIP-Seq) to analyze B cell-specific chromosomal conformation. We focused on the Pax5 gene that encodes a master regulator of B cell-lineage commitment and succeeded in identifying genomic regions that interact commonly with the mouse and human Pax5 gene by in vitro enChIP-Seq. In addition, we analyzed histone marks on identified interacting genomic regions by ChIP-Seq and deleted those regions by genome editing to analyze physiological importance of the genomic interactions for Pax5 expression and B cell homeostasis.
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| Free Research Field |
分子生物学
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| Academic Significance and Societal Importance of the Research Achievements |
本研究成果は、in vitro enChIP-Seq法が、特定遺伝子座を中心とした染色体三次元構造の解析に有用な技術であることを示している。in vitro enChIP-Seq法は、高度なバイオインフォマティクス処理を必要せずにゲノム領域間相互作用が解析できる技術であることから、3C法やその派生法と比較して汎用性が高い技術と考える。Pax5遺伝子座の転座が、B細胞リンパ腫に関連することが報告されていることから、本研究成果は、B細胞特異的な染色体三次元構造の生理的意義の解明のみならず、染色体三次元構造と転座の関連性の解明につながる可能性も秘めている。
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