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通过高通量 RNA 测序分析有或无新城疫病毒 LaSota 疫苗注射的中国鹧鸪胫骨组织的转录组。

Transcriptome analysis of thymic tissues from Chinese Partridge Shank chickens with or without Newcastle disease virus LaSota vaccine injection via high-throughput RNA sequencing.

机构信息

Henan University of Animal Husbandry and Economy, Zhengzhou, China.

Henan Chenxia Biomedical Co., Ltd, Zhengzhou, China.

出版信息

Bioengineered. 2022 Apr;13(4):9131-9144. doi: 10.1080/21655979.2021.2008737.

Abstract

The LaSota strain of Newcastle disease virus (NDV) is a commonly used vaccine to control Newcastle disease. However, improper immunization is a common reason for vaccine failure. Hence, it is imperative to thoroughly explore innate immunity-related molecular regulatory responses to the LaSota vaccine. In this text, 140 long non-coding RNAs (lncRNAs), 8 microRNAs (miRNAs), and 1514 mRNAs were identified to be differentially expressed by RNA sequencing analysis in the thymic tissues of Chinese Partridge Shank chickens after LaSota vaccine inoculation. Moreover, 70 dysregulated genes related to innate immunity were identified based on GO, Reactome pathway, and InnateDB annotations and differential expression analysis. Additionally, dysregulated lncRNAs and innate immunity-related mRNAs that could interact with dysregulated miRNAs were identified based on bioinformatics prediction analysis via the miRanda software and differential expression analysis. Among these transcripts, expression patterns of five lncRNAs, seven miRNAs, and six mRNAs were further examined by RT-qPCR assay. Both RNA-seq and RT-qPCR outcomes showed that 10 transcripts (MSTRG.22689.1, ENSGALT00000065826, ENSGALT00000059336, ENSGALT00000060887, gga-miR-6575-5p, gga-miR-6631-5p, gga-miR-1727, paraoxonase 2 (PON2), mitogen-activated protein kinase 10, and cystic fibrosis transmembrane conductance regulator (CFTR) were highly expressed, and 4 transcripts (MSTRG.188121.10, gga-miR-6655-5p, gga-miR-6548-3p, and matrix metallopeptidase 9 (MMP9) were low expressed after NDV infection. Additionally, two potential competing endogenous RNA networks (ENSGALT00000060887/gga-miR-6575-5p/PON2 or MSTRG.188121.10/gga-miR-6631-5p/MMP9) and some co-expression axes (ENSGALT00000065826/gga-miR-6631-5p, MSTRG.188121.10/gga-miR-6575-5p, MSTRG.188121.10/CFTR, ENSGALT00000060887/MMP9) were identified based on RT-qPCR and co-expression analyses. In conclusion, we identified multiple dysregulated lncRNAs, miRNAs, and mRNAs after LaSota infection and some potential regulatory networks for these dysregulated transcripts.

摘要

新城疫病毒(NDV)的 LaSota 株是一种常用于控制新城疫的疫苗。然而,免疫不当是疫苗失败的常见原因。因此,彻底探讨 LaSota 疫苗接种后与固有免疫相关的分子调控反应至关重要。在本文中,通过 RNA 测序分析,在接种 LaSota 疫苗后,中国鹧鸪胸肌组织中鉴定出 140 个长非编码 RNA(lncRNA)、8 个 microRNA(miRNA)和 1514 个 mRNA 的差异表达。此外,根据 GO、Reactome 通路和 InnateDB 注释和差异表达分析,鉴定出与固有免疫相关的 70 个失调基因。此外,通过 miRanda 软件的生物信息学预测分析和差异表达分析,鉴定出与失调 miRNA 相互作用的失调 lncRNA 和固有免疫相关的 mRNAs。在这些转录物中,通过 RT-qPCR 检测进一步检测了 5 个 lncRNA、7 个 miRNA 和 6 个 mRNA 的表达模式。RNA-seq 和 RT-qPCR 的结果均表明,10 个转录物(MSTRG.22689.1、ENSGALT00000065826、ENSGALT00000059336、ENSGALT00000060887、gga-miR-6575-5p、gga-miR-6631-5p、gga-miR-1727、paraoxonase 2(PON2)、mitogen-activated protein kinase 10 和囊性纤维化跨膜电导调节剂(CFTR)高度表达,而 4 个转录物(MSTRG.188121.10、gga-miR-6655-5p、gga-miR-6548-3p 和基质金属蛋白酶 9(MMP9)在 NDV 感染后表达较低。此外,根据 RT-qPCR 和共表达分析,还鉴定出两个潜在的竞争性内源 RNA 网络(ENSGALT00000060887/gga-miR-6575-5p/PON2 或 MSTRG.188121.10/gga-miR-6631-5p/MMP9)和一些共表达轴(ENSGALT00000065826/gga-miR-6631-5p、MSTRG.188121.10/gga-miR-6575-5p、MSTRG.188121.10/CFTR、ENSGALT00000060887/MMP9)。总之,我们在 LaSota 感染后鉴定出多个失调的 lncRNA、miRNA 和 mRNA,以及这些失调转录物的一些潜在调节网络。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bea6/9161911/a4327186e529/KBIE_A_2008737_F0001_OC.jpg

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