RNA sequencing-based analysis of the visceral tissue transcriptome of chicken embryo infected with F48E9 or La Sota
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ABSTRACT: We attempted to characterize the transcriptome of the chicken embryo during Newcastle disease virus (NDV) infection using RNA-sequencing analysis. The cDNAs derived from Total RNA of the pooled visceral tissues infected with F48E9 or La Sota were sequenced and analysed. The collected clean reads covered about 4.02% (2,341,868 reads) of the entire F48E8 reference sequence, while only 0.02% reads (13,886) were mapped to the La Sota genome. RNA-Seq datasets from groups La Sota, F48E9 and control, were respectively mapped to 71.76%, 68.55% and 70.05% of the reference genome Galgal 4.73. Compared with the control, 2,035 and 1,604 differentially expressed genes of hosts were found responding to F48E9 and La Sota infection, respectively. GO and KEGG pathway enriched various signalling pathways with elements playing roles in enhancing or preventing viral infection, like IFP35, NMI, Mx, OAS*A, IFITM5, STAT1 and IFNβ. So far, we know that velogenic NDV made far more transcripts during infection and caused significant impact on the host, showing a large number of genes in various pathways at high levels of expression.
Project description:Three cDNA libraries of mixed visceral tissues from F48E9, La Sota, or uninfected chicken embryos were constructed, and small-RNA deep sequencing was conducted to detect the expression levels of small-RNAs. Intergroup comparisons were used to identify changes in miRNA expression caused by NDV infection. La Sota affected the expression of 61 miRNAs (36 upregulated and 25 downregulated) at 36 hpi, and F48E9 infection altered the expression levels of 66 miRNAs (33 upregulated and 31 downregulated).
Project description:Newcastle disease virus (NDV) has emerged as an oncolytic agent in several cancers. Previous study has shown that NDV exerts cytolytic activity in glioma, however, the underlying mechanism has not been fully uncovered. Here the cytolytic activity of NDV in glioma and the associated mechanisms have been demonstrated. Infection with NDV inhibits cell proliferation and promotes cell apoptosis in LN229 cells. Further investigation showed that cytoplasmic organelle damage and cytoplasmic vacuolation were observed in LN229 cells after NDV infection. JC-1 staining assay proved that NDV caused cell apoptosis of LN229 cells by inducing mitochondrial dysfunction. We next speculated that NDV caused LN229 cells death through inducing necroptosis, but not ferroptosis, since the Fe2+ level did not alter after NDV infection. Furthermore, the NDV-caused cell death in LN229 cells was blocked by necroptosis inhibitor Nec1. Besides, RNA-seq analysis identified the different expression genes in NDV-infected LN229 cells. OASL, an antiviral gene, has been found to be directly induced by NDV infection. We also found that knockdown of OASL enhanced NDV infection-induced LN229 cells necroptosis. In summary, two aspects about cytolytic activity of NDV in glioma have been demonstrated. NDV presented cytolytic activity in glioma cells through inducing necroptosis. Additionally, targeting OASL may provide new strategy for enhancing necroptosis of glioma cells after NDV infection.
Project description:Newcastle disease (ND) affects a few hundred avian species including chicken, and the clinical outcome of Newcastle disease virus (NDV) infection ranges from mild to severe fatal disease depending on the NDV pathotype and the host species involved. Japanese quails serve as natural reservoirs of NDV and play important role in NDV epidemiology. While infection of chicken with velogenic NDV results in severe often fatal illness, the same infection in Japanese quails is results in in apparent infection. The molecular basis of this contrasting clinical outcomes of NDV infection is not yet known. We compared global gene expression in spleens of chicken and Japanese quails infected with a lentogenic or velogenic NDVs. We found contrasting regulation of key genes associated with NF-κB pathway and T-cell activation between chicken and Japanese quails. Our data suggests association of NDV resistance in Japanese quails to activation of NF-κB pathway and T cell proliferation.
Project description:The NDV GM strain was used to infect DEF cells with 1moi, while an uninfected group was set up as a control. Changes in transcript levels of different genes after NDV infection of duck cells were identified by high-throughput sequencing. It is hoped to reveal the unique antiviral mechanism of waterfowl in resisting NDV infection.
Project description:Newcastle disease virus (NDV) is an avian paramyxovirus that causes major economic losses to the poultry industry around the world, with NDV pathogenicity varying due to strain virulence differences. However, the impact of intracellular viral replication and the heterogeneity of host responses among cell types are unknown. Here, we investigated the heterogeneity of lung tissue cells in response to NDV infection in vivo and that of chicken embryo fibroblast cell line DF-1 to NDV infection in vitro using single-cell RNA sequencing.
Project description:N6-methyladenosine (m6A) modification is the most prevalent post-transcriptional modification of mRNA in eukaryotes and has been reported to play an important role in viral infection. However, the role of m6A modification during Newcastle disease virus infection (NDV) is not clear. In this study, we profiled the transcriptome-wide m6A modification of mRNA in NDV-infected chicken macrophages using MeRIP-seq. we identified a total of 9496 significantly changed peaks, of which 7015 peaks distributed across 3320 genes were significantly up-regulated and 2481 peaks distributed across 1264 genes were significantly down-regulated. Combined analysis of m6A peaks and mRNA expression revealed 1234 mRNAs with significantly altered methylation and expression levels after NDV infection, and m6A modification tended to have a negatively correlation with mRNA expression, suggesting that m6A modification may regulate the process of NDV infection by regulating gene expression, especially innate immunity-related genes. To our knowledge, This is the first comprehensive characterization of m6A patterns of the mRNA in chicken macrophages after NDV infection, and provides a valuable basis for further exploring the role of m6A modification in the process of NDV infection.
Project description:Purpose: To characterize the differential mRNA expression profiles of lung tissues upon PRRSV infection in different pig breeds, using NGS techonology, we sequenced mRNAs of the lungs of Tongcheng and Landrace pigs before (0 dpi) and after (3, 5, 7 dpi) infection with high-pathogenic PRRSV (HP-PRRSV). Methods: The mRNA expression profiles of the lungs of Tongcheng and Landrace pigs before (0 dpi) and after (3, 5, 7 dpi) HP-PRRSV infection were produced by using solexa platform. The raw reads with low qualities were filtered and the clean high quality reads were mapped to Ensembl Sus reference genome 10.2.71 using BOWTIE2. The unique mapped reads were retained for mRNA expression analysis. The raw reads counts of each mRNA were calculated by HTseq and the differentially expressed mRNA (Fold change >2; FDR <0.05) were called using DEGseq.
Project description:Genotyping based on genomic comparative hybridization of different isolates of coxiella burnetii compared to NMI reference strain Two-condition experiment, NMI vs. isolates. One replicate per isolate.