| 研究課題/領域番号 |
18K07966
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| 研究機関 | 金沢大学 |
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研究代表者 |
李 影奕 金沢大学, 医学系, 博士研究員 (70401940)
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| 研究分担者 |
本多 政夫 金沢大学, 保健学系, 教授 (00272980)
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| 研究期間 (年度) |
2018-04-01 – 2021-03-31
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| キーワード | Hepatitis B virus / infection / cccDNA / Dock11 |
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| 研究実績の概要 |
Hepatitis B virus (HBV) infection remains a major health problem worldwide. One major hurdle to treat HBV infection is the persistence of viral covalently closed circular DNA (cccDNA) that is not directly targeted by current anti-HBV therapeutics. Thus, it is imperative to understand the molecular mechanism that control cccDNA biogenesis, homeostasis and decay.
Until now, we identified DOCK11, a cccDNA maintenance gene, using our established cell culture model. The purpose of this project is to understand the roles of DOCK11 in cccDNA maintenance and subsequently clarify the possibility of DOCK11 as a target molecule for treating chronic HBV infection.
In this year, we have performed in vitro experiment and realized that HBV DNA could entry to the nucleus by Dock11 regulating X gene.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The project is going smoothly according to the experimental plan. The biological effects and mechanism of DOCK11 regarding HBV DNA maintenance were investigated and HBV Rc-DNA entering to the nucleus by Dock11 regulating X gene were clarified in vitro.
1) The overexpression and knockdown DOCK11 plasmids were constructed in lentiviral vector. 2) Stably DOCK11-expressing HepG2-NTCP-C4 cells were established in Dox-inducing system. 3) Stably DOCK11-knocked down HepG2-NTCP-C4 cells and Huh7 cells were established using CRISPR/Cas9 system. 4) The level of HBV DNA and cccDNA were increased or decreased by overexpression of Dock11 or silencing Dock11 by real-time PCR, Southern blotting, and Western blotting. 5) We demonstrated that HBV Rc-DNA entry to the nucleus by Dock11 regulating X gene.
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| 今後の研究の推進方策 |
We will perform the experiments according to the research plan.
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| 次年度使用額が生じた理由 |
There was a some difference in delivery prices for the experimental consumables.
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