A Genetic Compensation Phenomenon and Global Gene Expression Changes in Sex-miR-2766-3p Knockout Strain of Spodoptera exigua Hübner (Lepidoptera: Noctuidae).
A Genetic Compensation Phenomenon and Global Gene Expression Changes in Sex-miR-2766-3p Knockout Strain of Spodoptera exigua Hübner (Lepidoptera: Noctuidae).
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DOI：
10.3390/insects13111075
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发表时间：
2022-11-21
期刊：
影响因子：
3
通讯作者：
Hu Z
中科院分区：
文献类型：
--
作者：
Zuo Y;Wang Z;Ren X;Pei Y;Aioub AAA;Hu Z
mirna是一种内源性的小分子非编码RNA，在发育、免疫和变态等多种生物过程中起着至关重要的调节作用。由于一个miRNA可以调控数百个靶标，因此几乎所有的基因都可能在一定程度上受到miRNA的调控。因此，miRNA敲除可能会出现明显的症状，如发育缺陷和胚胎死亡。在本研究中，我们报道了S. exigua中Sex-miR-2766-3p敲除是非致死的，证实了miRNAs与其他miRNAs或其他途径冗余作用的理论。MicroRNAs （miRNAs）驱动靶mrna的转录后抑制，并在多种生物过程中发挥重要作用。miR-2766-3p在鳞翅目物种中是保守且丰富的，可能参与多种生物活动。在本研究中，Sex-miR-2766-3p被预测可能与夜蛾cap ' n ' collar isoform C （CncC）的3 ' untransledregion （UTR）结合，并通过荧光素酶报告系统筛选证实了Sex-miR-2766-3p调节SeCncC的表达。尽管CRISPR/Cas9已被广泛用于检测昆虫基因功能，但对miRNA功能的研究仍然相对少见。因此，我们使用CRISPR/Cas9敲除S. exigua的Sex-miR-2766-3p。然而，与WHS菌株相比，敲除菌株（2766-KO）中SeCncC的表达没有显著变化。这一结果表明，miRNA敲除可能由于遗传稳健性而缺乏表型。此外，我们使用转录组分析来检查Sex-miR-2766-3p敲除菌株的全局基因表达模式如何变化。RNA-seq数据显示，2766-KO菌株中有1746个差异表达基因（deg）上调，2183个差异表达基因（deg）下调，这可能是Sex-miR-2766-3p缺失或DNA损伤触发转录适应的结果。氧化石墨烯功能分类和KEGG通路分析表明，这些氧化石墨烯具有丰富的结合、催化活性、代谢过程和信号转导相关术语。我们的研究结果表明，S. exigua可以通过其他途径维持SeCncC的表达来弥补缺失的Sex-miR-2766-3p。
miRNAs are endogenous, small, non-coding RNA species that can be crucial regulators in a variety of biological processes, such as development, immunity, and metamorphosis. Because one miRNA may regulate hundreds of targets, nearly all genes are expected to be regulated by miRNAs to some degree. Since this is the case, miRNA knockouts may show obvious symptoms, such as developmental defects and embryonic mortality. In this study, we reported that Sex-miR-2766-3p knockout in S. exigua was non-lethal, confirming the theory that miRNAs function redundantly with other miRNAs or other pathways. MicroRNAs (miRNAs) drive the post-transcriptional repression of target mRNAs and play important roles in a variety of biological processes. miR-2766-3p is conserved and abundant in Lepidopteran species and may be involved in a variety of biological activities. In this study, Sex-miR-2766-3p was predicted to potentially bind to the 3′ untranslated region (UTR) of cap ‘n’ collar isoform C (CncC) in Spodoptera exigua, and Sex-miR-2766-3p was confirmed to regulate the expression of SeCncC through screening with a luciferase reporter system. Although CRISPR/Cas9 has been extensively utilized to examine insect gene function, studies of miRNA function are still relatively uncommon. Thus, we employed CRISPR/Cas9 to knock out Sex-miR-2766-3p from S. exigua. However, the expression of SeCncC was not significantly altered in the knockout strain (2766-KO) compared with that of the WHS strain. This result suggested that a miRNA knockout might lack phenotypes because of genetic robustness. Additionally, we used transcriptome analysis to examine how the global gene expression patterns of the Sex-miR-2766-3p knockout strain varied. RNA-seq data revealed 1746 upregulated and 2183 downregulated differentially expressed genes (DEGs) in the 2766-KO strain, which might be the result of Sex-miR-2766-3p loss or DNA lesions as the trigger for transcriptional adaptation. GO function classification and KEGG pathway analyses showed that these DEGs were enriched for terms related to binding, catalytic activity, metabolic process, and signal transduction. Our findings demonstrated that S. exigua could compensate for the missing Sex-miR-2766-3p by maintaining the expression of SeCncC by other pathways.
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