2022 Fiscal Year Final Research Report
Precise regulation of SOX9 expression during gonadal development by the testis specific enhancers using a genetically humanized mouse model
Project/Area Number |
20K18208
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Research Category |
Grant-in-Aid for Early-Career Scientists
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Allocation Type | Multi-year Fund |
Review Section |
Basic Section 56040:Obstetrics and gynecology-related
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Research Institution | National Center for Child Health and Development |
Principal Investigator |
Tsuji-Hosokawa Atsumi 国立研究開発法人国立成育医療研究センター, システム発生・再生医学研究部, (非)研究員 (40842414)
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Project Period (FY) |
2020-04-01 – 2023-03-31
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Keywords | Sox9 / sex determination / testis / enhancer / regulatory region |
Outline of Final Research Achievements |
SOX9/Sox9 is a master gene for testis development. This study aimed to analyze function of candidate sequence for human SOX9 enhancer, hXYSRa, in in vivo setting. 1) in vitro reporter assay utilizing luciferase expression plasmid revealed enhancer function of hXYSRa. We identified one of transcriptional factor binding motifs within the sequence. 2) We generated genetically humanized mice of Sox9 enhancer. The homologous Sox9 enhancer region in murine genome was replaced with hXYSRa sequence (hXYSRa sub mice) and development of reproductive system in the model mice was screened. Some of hXYSRa sub/sub mice with XY karyotype showed virilization of external/internal genitalia when the line was backcrossed to the DBA/2 strain and overexpressed human SRY. This result suggested regulatory regions in human genome can compensate homologous sequence in murine genome and can be analyzed in the animal model.
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Free Research Field |
genome editing
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Academic Significance and Societal Importance of the Research Achievements |
本研究では、ヒトSOX9エンハンサー候補領域について関連転写因子の結合部位を明らかにし、生殖腺分化に重要な遺伝子の発現調節解明に寄与した。また発現調節領域について、マウスゲノムの相同領域とヒト相同配列を置換した場合、一定条件においてはヒトエンハンサーをモデル生物内で機能解析できる可能性が示唆され、non coding regionの機能解析において重要な知見をもたらしたと言える。
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