Molecular cloning, expression and mimicking antiviral activity analysis of retinoic acid-inducible gene-I in duck (Anas platyrhynchos)
Molecular cloning, expression and mimicking antiviral activity analysis of retinoic acid-inducible gene-I in duck (Anas platyrhynchos)
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鸭视黄酸诱导基因-I的分子克隆、表达及模拟抗病毒活性分析
DOI：
10.1007/s12041-020-1187-x
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发表时间：
2020
影响因子：
1.5
通讯作者：
Lu Lizhi
中科院分区：
文献类型：
--
作者：
Gu Tiantian;Li Guoqin;Tian Yong;Chen Li;Wu Xinsheng;Zeng Tao;Xu Qi;Vladyslav Spyrydonov;Chen Guohong;Lu Lizhi
细胞内双链 RNA (dsRNA) 是许多病毒复制的主要标志。先天免疫系统的模式识别受体 (PRR) 检测到 dsRNA 并启动抗病毒反应。视黄酸诱导基因 I (RIG-I) 是 PRR 的成员，在 dsRNA 诱导的信号传导中发挥重要的调节作用。在本研究中，利用逆转录聚合酶链反应（RT-PCR）和cDNA末端快速扩增（RACE）技术克隆了鸭RIG-I（duRIG-I）的全长互补DNA（cDNA）。 duRIG-I的cDNA包含97-bp 5'UTR、141-bp 3'-UTR和2802bp完整开放阅读框(ORF)，编码933个氨基酸。多重序列比对表明duRIG-I与其他脊椎动物的RIG-I具有高度相似性。实时定量PCR（qRT-PCR）分析显示，duRIG-I mRNA在所有测试组织中均有表达，其中肝脏、心脏、脾脏、肾脏和胸腺中表达量较高，而十二指肠中表达量较低。 duRIG-I 可以通过聚 (I:C) 处理来诱导。此外，duRIG-I的过表达显着激活了poly(I:C)诱导的IFN-β、IRF7、TRIF、Mx、STAT1和STAT2 mRNA的转录，而duRIG-I敲低则显示出相反的结果。总的来说，我们的结果表明 duRIG-I 可能是模拟鸭体内抗病毒状态的重要受体，这需要进一步的研究来证明可能的机制。
Intracellular double-stranded RNA (dsRNA) is a chief sign of replication for many viruses. Pattern recognition receptors (PRRs) of the innate immune system detected the dsRNA and initiate the antiviral responses. Retinoic acid-inducible gene I (RIG-I), a member of PRRs, plays an essential regulatory role in dsRNA-induced signalling. In this study, the full-length complementary DNA (cDNA) of duck RIG-I (duRIG-I) was cloned using the reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of the cDNA ends (RACE). The cDNA of duRIG-I contained 97-bp 5 ' UTR, 141-bp 3 '-UTR and 2802 bp complete open-reading frame (ORF) encoding 933 amino acids. Multiple sequence alignments showed that duRIG-I shared high similarity with RIG-I from other vertebrates. Quantitative real-time PCR (qRT-PCR) analysis revealed that duRIG-I mRNA was expressed in all tested tissues, with high levels in the liver, heart, spleen, kidney and thymus, while lower in the duodenum. duRIG-I could be induced by treatment with poly(I:C). Further, overexpression of duRIG-I significantly activated the transcription of poly(I:C)-induced IFN-beta, IRF7, TRIF, Mx, STAT1 and STAT2 mRNA, and duRIG-I knockdown showed the opposite results. Overall, our results suggested that duRIG-I could be an important receptor for mimicking antiviral state in duck, which warrant further studies to show the possible mechanism.