Vector Borne Disease Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi, India.
Department of Biochemistry, Faculty of Science, Jamia Hamdard, New Delhi, India.
Sci Rep. 2017 Dec 15;7(1):17666. doi: 10.1038/s41598-017-18043-0.
Arboviruses that replicate in mosquitoes activate innate immune response within mosquitoes. Regulatory non-coding microRNAs (miRNA) are known to be modulated in mosquitoes during chikungunya infection. However, information about targets of these miRNAs is scant. The present study was aimed to identify and analyze targets of miRNAs that are regulated during chikungunya virus (CHIKV) replication in Aedes aegypti cells and in the mosquito. Employing next-generation sequencing technologies, we identified a total of 126 miRNAs from the Ae. aegypti cell line Aag2. Of these, 13 miRNAs were found to be regulated during CHIKV infection. Putative targets of three of the most significantly regulated miRNAs- miR-100, miR-2b and miR-989 were also analyzed using quantitative PCRs, in cell lines and in mosquitoes, to validate whether they were the targets of the miRNAs. Our study expanded the list of miRNAs known in Ae. aegypti and predicted targets for the significantly regulated miRNAs. Further analysis of some of these targets revealed that ubiquitin-related modifier is a target of miRNA miR-2b and plays a significant role in chikungunya replication.
在蚊子中复制的虫媒病毒会激活蚊子的先天免疫反应。在基孔肯雅热感染期间，已知调节性非编码 microRNA（miRNA）在蚊子中被调节。然而，关于这些 miRNA 靶标的信息很少。本研究旨在鉴定和分析在埃及伊蚊细胞和蚊子中复制的基孔肯雅病毒（CHIKV）过程中调节的 miRNA 的靶标。本研究采用下一代测序技术，从埃及伊蚊细胞系 Aag2 中总共鉴定出 126 种 miRNA。其中，有 13 种 miRNA 在 CHIKV 感染过程中被发现受到调节。使用定量 PCR 技术，在细胞系和蚊子中分析了三个调节最显著的 miRNA（miR-100、miR-2b 和 miR-989）的假定靶标，以验证它们是否为 miRNA 的靶标。本研究扩展了已知在埃及伊蚊中存在的 miRNA 列表，并预测了显著调节的 miRNA 的靶标。对其中一些靶标的进一步分析表明，泛素相关修饰物是 miRNA miR-2b 的靶标，并在基孔肯雅热复制中发挥重要作用。
