College of Plant Protection, Yangzhou University, Yangzhou 225009, China.
Laboratory for Comparative Genomics and Bioinformatics & Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Science, Nanjing Normal University, Nanjing 210046, China.
Genes (Basel). 2024 May 8;15(5):601. doi: 10.3390/genes15050601.
Innate immune response is the first line of host defense against pathogenic microorganisms, and its excessive or insufficient activation is detrimental to the organism. Many individual microRNAs (miRNAs) have emerged as crucial post-transcriptional regulators of immune homeostasis in . However, the synergistical regulation of miRNAs located within a cluster on the Imd-immune pathway remains obscured. In our study, a genetic screening with 52 transgenic UAS-miRNAs was performed to identify ten miRNAs or miRNA clusters, including the miR310313 cluster, which may function on Imd-dependent immune responses. The miRNA RT-qPCR analysis showed that the expression of miR-310313 cluster members exhibited an increase at 6-12 h post infection. Furthermore, the overexpression of the miR-310313 cluster impaired the survival. And the overexpression of miR-310/311/312 reduced expression, an indication of Imd pathway induced by Gram-negative bacteria. Conversely, the knockdown of miR-310/311/312 led to increases in expression. The Luciferase reporter expression assays and RT-qPCR analysis confirmed that miR-310313 cluster members directly co-targeted and inhibited transcription. These findings reveal that the members of the miR-310~313 cluster synergistically inhibit Imd-dependent immune responses by co-targeting the Imd gene in .
先天免疫反应是宿主防御致病微生物的第一道防线，其过度或不足激活都对机体有害。许多个体 microRNAs（miRNAs）已成为 中免疫稳态的关键转录后调节因子。然而，位于 Imd-免疫途径上的 miRNA 簇的协同调节仍然不清楚。在我们的研究中，进行了 52 种转基因 UAS-miRNAs 的遗传筛选，以鉴定十个可能在 Imd 依赖的免疫反应中起作用的 miRNAs 或 miRNA 簇，包括 miR310313 簇。miRNA RT-qPCR 分析显示，miR-310313 簇成员的表达在 感染后 6-12 小时增加。此外，miR-310313 簇的过表达会损害 的存活。miR-310/311/312 的过表达降低了 的表达，这表明革兰氏阴性菌诱导了 Imd 途径。相反，miR-310/311/312 的敲低导致 的表达增加。荧光素酶报告表达测定和 RT-qPCR 分析证实，miR-310313 簇成员通过直接靶向和抑制 Imd 基因的转录来协同抑制 Imd 依赖的免疫反应。这些发现表明，miR-310~313 簇成员通过共同靶向 中的 Imd 基因，协同抑制 Imd 依赖的免疫反应。
