Project description:Pseudorabies, an acute infectious disease caused by pseudorabies virus (PRV), has caused enormous economic losses to the breeding industry in many countries worldwide. Accumulating evidence indicates that long non-coding RNAs (lncRNAs) may play important roles in the antiviral responses. However, little is known about the identification and functions of swine lncRNAs in cellular antiviral responses against PRV II. In this study, we detected the expression profiles of host lncRNAs and mRNAs from PRV-DX, a wild-type (WT) strain of PRV II, and its attenuated gE-TK- PRV-DX infected cells by high-throughput RNA sequencing. Finally, we identified differentially expressed (DE) 664 lncRNAs and DE 7,199 mRNAs from PRV-DX infected cells, and 654 DE lncRNAs and DE 7,149 mRNAs from gE-TK- PRV-DX infected cells when compared to MOCK infected cells, respectively, especially including 469 common DE lncRNAs and 5836 DE mRNAs. Besides 276 DE lncRNAs and 2,272 DE mRNAs between PRV-DX and gE-TK- PRV-DX infected cells were identified. The potential functions of the significant DE lncRNAs were involved in interleukin secretion, axon extension and metabolic process based on the Gene ontology and Kyoto Encyclopedia of Genes and Genomes databases. Taken together, results highlighted the potentials of lncRNA as targets for antiviral therapy and enriched the current knowledge of the mechanisms underlying the interaction between PRV II and its host cells.
Project description:Circular RNAs (circRNAs) and microRNAs (miRNAs) participate in regulating many biological processes. However, their roles in PrV-II pathogenicity are largely unknown. Here, we analyzed the expression profile of circRNAs and miRNAs in the PrV-DX, a wild-type (WT) strain of PRV-II, and its attenuated gE-TK- PRV-DX infected cells by high-throughput sequencing.
Project description:Purpose: Evaluation of the m6A modification of PRV and PK15 transcripts during PRV infection Methods: Porcine kidney cell line PK15 was uninfected or infected with PRV for 24 hours. Total RNA from each sample were extracted. Intact mRNA was isolated from total RNA samples and then chemically fragmented to 300-nucleoside-long fragments. Fragmented mRNAs were immunoprecipitated with anti-N6-methyadenosine (m6A) antibody (a part of the fragmented mRNAs was kept as input). Both m6A enriched mRNAs and input mRNAs were concentrated for RNA-seq libraries construction. The libraries were forwarded to sequencing run on Illumina NovaSeq 6000. Results: PRV transcripts were m6A modified during PRV infection and PRV infection changed m6A modification profiles of PK15 transcripts.
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI). Four time points were studied: 1h, 2h, 4h and 8h post-I and MI. Four replicates of I and MI were analysed. Keywords: Pig, PrV, Pk15 cells, kinetics
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI).Six time points were studied: just after I and MI, 1h, 2h, 4h, 8h and 12h post-I and MI. For this study, a pig DNA/cDNA microarray containing genes of the SLA region, additional genes encoding other important immunological molecules and all the PrV genes was constructed. Keywords: infection time course
Project description:To study in vitro the epithelial cells and PrV interactions during infection, we followed PrV and PK15 cells transcriptome modifications during time-course infection (I) and mock-infection (MI). Four time points were studied: 1h, 2h, 4h and 8h post-I and MI. Four replicates of I and MI were analysed. 32 samples - The hybridization scheme which can be define as dye-switch was chosen. A balanced loop design with two independent loops, each loop containing 2 replicates of PrV infection and MI, was used. In total, 32 slides were used in this experiment.