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全基因组整合分析 miRNA 和 mRNA 表达谱,以鉴定对经典猪瘟疫苗病毒反应的差异表达 miR-22-5p 和 miR-27b-5p。

Genome-wide integrated analysis of miRNA and mRNA expression profiles to identify differentially expressed miR-22-5p and miR-27b-5p in response to classical swine fever vaccine virus.

机构信息

Animal Genetics, Indian Veterinary Research Institute, Izatnagar, Bareilly, 143122, India.

Standardization Division, Indian Veterinary Research Institute, Izatnagar, Bareilly, 143122, India.

出版信息

Funct Integr Genomics. 2019 Nov;19(6):901-918. doi: 10.1007/s10142-019-00689-w. Epub 2019 May 28.

Abstract

The present study was conducted to identify the differentially expressed miRNAs (DE miRNAs) in the peripheral blood mononuclear cells of crossbred pigs in response to CSF vaccination on 7 and 21 days of post vaccination as compared to unvaccinated control (0 dpv). Simultaneously, set of miRNA was predicted using mRNA seq data at same time point. The proportion of CD4CD8 and CD4CD8 increased after vaccination, and the mean percentage inhibition was 86.89% at 21 dpv. It was observed that 22 miRNAs were commonly expressed on both the time points. Out of predicted DE miRNAs, it was found that 40 and 35 DE miRNAs were common, obtained from miRNA seq analysis and predicted using mRNA seq data on 7 dpv versus 0 dpv and 21 dpv versus 0 dpv respectively. Two DE miRNAs, ssc-miR-22-5p and ssc-miR-27b-5p, were selected based on their log fold change and functions of their target genes in immune process/pathway of viral infections. The validations of DE miRNAs using qRT-PCR were in concordance with miRNA seq analysis. Two set of target genes, CD40 and SWAP70 (target gene of ssc-miR-22-5p) and TLR4 and Lyn (target gene of ssc-miR-27b-5p), were validated and were in concordance with results of RNA seq analysis at a particular time point (except TLR4). The first report of genome-wide identification of differentially expressed miRNA in response to live attenuated vaccine virus of classical swine fever revealed miR-22-5p and miR-27b-5p were differentially expressed at 7 dpv and 21 dpv.

摘要

本研究旨在鉴定 CSF 疫苗接种后 7 天和 21 天杂交猪外周血单个核细胞中差异表达的 miRNAs(DE miRNAs)与未接种对照(0 dpv)相比。同时,在同一时间点使用 mRNA seq 数据预测了一组 miRNA。疫苗接种后 CD4CD8 和 CD4CD8 的比例增加,21 dpv 的平均抑制率为 86.89%。观察到 22 个 miRNA 在两个时间点均有表达。在预测的 DE miRNAs 中,发现 miRNA seq 分析和使用 mRNA seq 数据在 7 dpv 与 0 dpv 以及 21 dpv 与 0 dpv 之间分别预测到 40 和 35 个 DE miRNAs 是共同的。根据其对数倍数变化和在免疫过程/病毒感染途径中其靶基因的功能,选择了两个差异表达 miRNA,ssc-miR-22-5p 和 ssc-miR-27b-5p。使用 qRT-PCR 对 DE miRNAs 的验证与 miRNA seq 分析一致。两组靶基因 CD40 和 SWAP70(ssc-miR-22-5p 的靶基因)和 TLR4 和 Lyn(ssc-miR-27b-5p 的靶基因)进行了验证,并与特定时间点的 RNA seq 分析结果一致(除 TLR4 外)。这是首次报道针对经典猪瘟活弱毒疫苗的全基因组差异表达 miRNA 鉴定,表明 miR-22-5p 和 miR-27b-5p 在 7 dpv 和 21 dpv 时差异表达。

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