Characterization of the biological functions of the neuronal transcription factor MIBP1 using genome editing
| Project/Area Number |
17K07245
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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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| Research Field |
Genome biology
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| Research Institution | Kinjo Gakuin University |
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Principal Investigator |
Tomoko Tahira 金城学院大学, 薬学部, 准教授 (50155230)
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| Project Period (FY) |
2017-04-01 – 2023-03-31
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| Project Status |
Completed (Fiscal Year 2022)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 転写因子 / ゲノム編集 / 遺伝子発現調節 |
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| Outline of Final Research Achievements |
The MIBP1 (HIVEP2) gene encodes a 270 kDa protein that acts as a transcription factor, by binding to specific DNA sequences. Loss of function mutations in the MIBP1 gene have been associated with intellectual disabilities, suggesting that MIBP1 is crucial for proper brain development and function.
To identify target genes for transcriptional regulation by MIBP1 protein, we tried to insert FLAG-tag sequence to endogenous MIBP1 gene by genome editing of various cell lines. One of established cell lines, M15, which is derived from HCT116 colon cancer cells have both wild type and edited allele. MIBP1 expression was induced immediately after TNF-alpha treatment from the edited allele as well as the wild type allele. Increased mRNA expression was accompanied by enhanced expression of FLAG-tagged MIBP1 protein. As a result of genome editing, changes in mRNA stability were observed.
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では、転写因子をコードする内在性MIBP1遺伝子に対してゲノム編集によりエピトープタグ配列を導入することで、タグ配列に対する抗体を用いて高感度にタンパク質の解析を行うことが可能となった。これは抗体の品質によってはタンパク質解析の結果の再現性が得られない問題を解決するものである。本研究により、MIBP1タンパク質の発現が炎症性サイトカインであるTNF-αにより上昇することが明らかになった。一方、ゲノム編集で本来なかった配列を挿入したことによりmRNAの安定性が変化するという副次的な影響もあることがわかった。
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Report
(7 results)
Research Products
(9 results)
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[Journal Article] Idarubicin, an Anthracycline, Induces Oxidative DNA Damage in the Presence of Copper (II).2020
Author(s)
Mizutani H, Shiga C, Imai M, Ikemura K, Kitamura Y, Ohta K, Miyazawa D, Sakanashi M, Tahira T, Maeda T, Hiraku Y, Kawanishi S.
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Journal Title
Anticancer Research
Volume: 40
Issue: 10
Pages: 5399-5404
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] 脳神経機能に関与するHIVEP2遺伝子の転写および転写後調節の解析2023
Author(s)
山本 蒼, 中川 朱里, 米田 幸香, 川原 侑奈, 須田 櫻, 髙田 真衣, 夏目 佳乃, 水谷 百合江, 山本 真菜, 宮澤 大介, 田平 知子
Organizer
日本薬学会第143年会
Related Report
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