Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore City, Singapore.
School of Biological Sciences, Nanyang Technological University, Singapore City, Singapore.
FEBS Lett. 2019 Nov;593(21):3003-3014. doi: 10.1002/1873-3468.13564. Epub 2019 Aug 19.
The cytoplasmic immune sensor RIG-I detects viral RNA and initiates an antiviral immune response upon activation. It has become a potential target for vaccination and immunotherapies. To develop the smallest but potent immunomodulatory RNA (immRNAs) species, we performed structure-guided RNA design and used biochemical, structural, and cell-based methods to select and characterize the immRNAs. We demonstrated that inserting guanosine at position 9 to the 10mer RNA hairpin (3p10LG9) activates RIG-I more robustly than the parental RNA. 3p10LG9 interacts strongly with the RIG-I helicase-CTD RNA sensing module and disrupts the auto-inhibitory interaction between the HEL2i and CARDs domains. We further showed that 3p10LA9 has a stronger cellular activity than 3p10LG9. Collectively, purine insertion at position 9 of the immRNA species triggered more robust activation of RIG-1.
细胞质免疫传感器 RIG-I 检测病毒 RNA，并在激活后启动抗病毒免疫反应。它已成为疫苗接种和免疫疗法的潜在目标。为了开发最小但有效的免疫调节 RNA（immRNAs），我们进行了结构导向的 RNA 设计，并使用生化、结构和基于细胞的方法来选择和表征 immRNAs。我们证明，在 10 mer RNA 发夹（3p10LG9）的第 9 位插入鸟嘌呤比亲本 RNA 更能强烈激活 RIG-I。3p10LG9 与 RIG-I 解旋酶-CTD RNA 感应模块强烈相互作用，并破坏 HEL2i 和 CARDs 结构域之间的自动抑制相互作用。我们还表明，3p10LA9 比 3p10LG9 具有更强的细胞活性。总之，immRNA 种的第 9 位嘌呤插入可触发 RIG-1 的更强烈激活。
