| 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)
|
|---|
| 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.
|
