A novel approach to inhibition of human virus infection : inhibition of DNA virus replication by artificial DNA-binding proteins
| Project/Area Number |
17550154
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Kyoto University |
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Principal Investigator |
TAKAHASHI Sera Kyoto University, Graduate School of Engineering, Associate Professor, 工学研究科, 助教授 (10362443)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | DNA virus / Inhibition of replication / DNA-binding protein |
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| Research Abstract |
Recently, we demonstrated that plant DNA virus replication was inhibited in planta by using an artificial zinc-finger protein (AZP) and created AZP-based transgenic plants resistant to DNA virus infection. Here we apply the AZP technology to inhibition of replication of a mammalian DNA virus. Two AZPs, designated AZP-1 and AZP-2, were designed using our nondegenerated recognition code table and constructed to block binding of a viral replication protein to its replication origin. Both the newly designed AZPs had much higher affinities towards the replication origin than the viral replication protein, and efficiently blocked the binding in vitro. Next, we established a transient replication assay to evaluate the ability of AZPs to inhibit virus replication. In the transient replication assays, both AZPs inhibited the viral DNA replication : AZP-2, especially, reduced the replication level to approximately 10%. We also demonstrated it in transient replication assays with mutant replication origins that AZP_<HPV>-2 could precisely recognize the replication origin in mammalian cells, thereby inhibiting virus replication. Thus, it was demonstrated that the AZP technology could be applied not only to plant DNA viruses, but also to mammalian DNA viruses.
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Report
(3 results)
Research Products
(6 results)
