Shimura Hanako, Fukagawa Takako, Meguro Ayano, Yamada Hirokazu, Oh-Hira Mahito, Sano Shinsuke, Masuta Chikara
Laboratory of Cell Biology and Manipulation, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
FEBS Lett. 2008 Dec 10;582(29):4047-52. doi: 10.1016/j.febslet.2008.10.046. Epub 2008 Nov 7.
To find out whether we can control plant virus diseases by blocking viral RNA silencing suppressors (RSSs), we developed a strategy to screen inhibitors that block the association of RSSs with siRNAs using a surface plasmon resonance assay. The screened chemicals were tested in competition with RSSs for binding to siRNAs using a mobility shift assay. We then confirmed that tested chemicals actually inhibited the RSS activity in vivo using a protoplast assay which was developed for this purpose. This entire system can be adapted to screening inhibitors of not only plant viruses but also some animal viruses possessing RSSs.
为了弄清楚我们是否能够通过阻断病毒RNA沉默抑制子(RSSs)来控制植物病毒病,我们开发了一种策略,利用表面等离子体共振分析筛选能够阻断RSSs与小干扰RNA(siRNAs)结合的抑制剂。使用迁移率变动分析对筛选出的化学物质与RSSs竞争结合siRNAs的能力进行测试。然后,我们利用为此目的开发的原生质体分析证实了测试的化学物质在体内确实抑制了RSS活性。整个系统不仅可以用于筛选植物病毒的抑制剂,也可用于筛选一些具有RSSs的动物病毒的抑制剂。