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MicroRNA profile analysis of feline kidney cell line infected with Feline Panleukopenia Virus using deep sequencing

ABSTRACT: Purpose:MicroRNAs (miRNAs) are members of a rapidly growing class of small endogenous non-coding RNAs that play crucial roles in post-transcriptional regulator of gene expression in many biological processes. Feline Panleukopenia Virus (FPV) is a highly infectious pathogen that causes severe disease in pets, economically important animals and wildlife in worldwide. However, the molecular mechanisms underlying the pathogenicity of FPV have not been completely clear. To study the involvement of miRNAs in the FPV infection process, miRNAs expression profiles were identified via deep sequencing in the feline kidney cell line (F81) infected and uninfected with FPV. Methods:miRNA-sequencing analysis was performed on an Illumina Hiseq 2500 (LC Sciences, USA) following the vendor's recommended protocol Results:As a result, 673 known miRNAs belonging to 210 families and 278 novel miRNAs were identified. Then we found 57 significantly differential expression miRNAs by comparing the results between uninfected and FPV-infected groups. Furthermore, stem-loop qRT-PCR was applied to validate and profile the expression of the randomly selected miRNAs; the results were consistent with those by deep sequencing. Furthermore, the potential target genes were predicted. The target genes of differential expression miRNAs were analyzed by GO and KEGG pathway. Conclusions:The identification of miRNAs in feline kidney cell line before and after infection with Feline Panleukopenia Virus will provide new information and enhance our understanding of the functions of miRNAs in regulating biological processes.

ORGANISM(S): Felis catus

PROVIDER: GSE122381 | GEO | 2020/12/20

REPOSITORIES: GEO

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Project description:Purporse: FPV can cause a viral disease that occurs in cats, causing severe leukopenia, gastro-enteritis and nervous signs and leads to significant losses. LncRNAs biochemically resemble mRNAs posited by Jacob and Monod, yet do not template protein synthesis. Rather, lncRNAs functioning as RNA genes to play an important role in many biological and pathological processes such as inflammation and viral infection. However, lncRNAs have not been comprehensively identified in felines. Here, microarrays of mRNAs and lncRNAs were globally performed to identify the lncRNAs related to immune response of feline panleukopenia virus infection in F81 cells. Methods:lncRNA and mRNA sequencing analysis was performed on an Illumina Hiseq 4000 (LC Sciences, USA) following the vendor's recommended protocol Results: Through RNA sequencing, we performed a comprehensive analysis of lncRNAs and mRNAs in FPV infected F81 cells. F81 cells infected or uninfected FPV were used to prepare six libraries to generate 14.66 to 15.45 gigabytes (GB) clean reads per sample. We obtained 90588 novel lncRNAs, and 291 are differential expressed lncRNAs between FPV infected cells and uninfected cells. Consistent with previous studies, lncRNAs showed features distinct from mRNAs, such as shorter lengths, lesser exon numbers, and lower expression levels compared to mRNAs. The GO enrichment and KEGG pathway analyses showed that a majority of genes and lncRNAs were mainly related to the immune response of host cells with FPV infection. By differentially expressed target genes as well as co-expression network construction analysis, 21 key differentially expressed genes (19 upregulated & 2 downregulated) related to immune regulation of FPV infection in F81 cells were identified. Moreover, fifteen differentially expressed genes (five lncRNAs, ten mRNAs) were selected and independently validated by reverse-transcription qPCR and the results were consistent with the sequencing results. Conclusion: The present study is the first to provide insights into the biological connection of lncRNAs and FPV infection, which can be further explore for the potential link between lncRNA expression and the pathogenesis of FPV infection.

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MicroRNA profile analysis of feline kidney cell line infected with Feline Panleukopenia Virus using deep sequencing

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