Effect of SRCAP on gene expression during PRRSV infection of Marc-145 cells.
Ontology highlight
ABSTRACT:
ORGANISM(S): Chlorocebus sabaeus
PROVIDER: GSE232483 | GEO | 2023/05/21
REPOSITORIES: GEO
Similar Datasets
Project description:Effect of SRCAP on gene expression during PRRSV infection of Marc-145 cells.
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Project description:Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). It clinically manifests as fever, bleeding, and respiratory symptoms in pigs of different age groups, as well as reproductive disorders, causing huge economic losses to the global swine industry. RNA interference (RNAi), a highly conserved mechanism mediating post-transcriptional gene silencing, exerts antiviral effects by cleaving viral double-stranded RNA to generate virus-derived siRNA (vsiRNA). VsiRNA-mediated antiviral immunity is widespread in plants and invertebrates, while in mammals, studies have focused on reducing PRRSV viral load in pigs via siRNA delivery. Therefore, PRRSV strains (LY and LY-R) were primarily selected as virus models to infect PAM and Marc-145 cells, respectively. Then, miRNA and vsiRNA analysis were performed and bioinformatics analysis revealed multiple differentially expressed miRNAs following PRRSV infection. Further enrichment analysis indicated that these miRNAs were mainly enriched in metabolic, endocytic, and phosphorylation signaling pathways. Additionally, in Marc-145 cells infected with PRRSV-LY, high abundance of vsiRNA was detected within the PRRSV-N genome. The N-vsiRNA was inserted into vector psilencer4.1. to construct a PS-N plasmid, which is capable of exerting anti-PRRSV (LY, LY-R, and NADC30-like) activity in Marc-145 cells. Finally, the PS-N plasmid was formulated with PLGA-PEI to prepare a nanoparticle mixture, which was delivered to pigs via intramuscular injection to alleviate tissue damage caused by PRRSV-LY infection to a certain extent. In summary, the porcine RNAi system and vsiRNA generated via viral cleavage play a critical role in inhibiting virus replication. This further demonstrates the feasibility of vsiRNA-based antiviral strategies and offers a reference for the prevention and control of various RNA viruses.
2025-10-01 | GSE308579 | GEO
Project description:We found that fuse ΔLMP1 to MAVS could strengthen MAVS mediated inhibition of PRRSV replication in MARC-145 cells. To better understand the biological function of the fusion protein ΔLMP1-MAVS, overall gene expression of MARC-145 cells transfected with ΔLMP1-MAVS or MAVS was evaluated by mRNA-seq. The result showed that ΔLMP1-MAVS upregulated a number of genes associated with innate immune responses to viral infection, including plenty of interferon-stimulated genes. This study provides reference date to research the working mechanism of ΔLMP1-MAVS.
2019-03-05 | GSE111530 | GEO
Project description:Porcine respiratory and reproductive syndrome virus (PRRSV) is a virus infecting swine and causes swine abortion. Previously, non-structural protein 11 (Nsp11) from PRRSV was shown to have inhibitory function to type I IFN signaling. In this project, we want to see in addition to type I IFN, whether other cellular pathways are influenced by Nsp11 systemtically. A cell line stably expressing PRRSV Nsp11 was established, designated as MARC-Nsp11 cells, and an RNA microarray was conducted using these cells and WT MARC-145 cells
2013-12-16 | GSE52335 | GEO
Project description:Porcine respiratory and reproductive syndrome virus (PRRSV) is a virus infecting swine and causes swine abortion. Previously, non-structural protein 11 (Nsp11) from PRRSV was shown to have inhibitory function to type I IFN signaling. In this project, we want to see in addition to type I IFN, whether other cellular pathways are influenced by Nsp11 systemtically. A cell line stably expressing PRRSV Nsp11 was established, designated as MARC-Nsp11 cells, and an RNA microarray was conducted using these cells and WT MARC-145 cells MARC-145 and MARC-Nsp11 cells were seeded one day prior to experiments and total cellular RNAs were extracted using Trizol (Invitrogen) and purified by RNeasy mini kit (Qiagen). The quantity and quality of RNA were determined using an Align 2100 bioanalyzer (Agilent Technologies, Palo, Alto, CA, USA), and the RNA integrity was determined above 7. The RNA samples were then subjected to microarray using Human Gene 1.0 ST arrays (Affymetrix UK Ltd, High Wycombe, UK) at the Keck Biotechnology Center, University of Illinois, Urbana, IL). The microarray was repeated twice in duplicates each.
2013-12-16 | E-GEOD-52335 | biostudies-arrayexpress
Project description:Virus-derived siRNA mediated inhibition of PRRSV replication in pigs [Marc-145]
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Project description:Cell lines derived from Chlorocebus sabaeus kidney were infected with an isolate of PRRSV-1 or PRRSV-2 and RNASeq was performed (this entry) in parallel with ribosome profiling (see related accession numbers). These RNASeq datasets provide information on the transcriptome of PRRSV-infected cells and were used to detect novel viral transcripts and perform differential gene expression analysis on host transcripts. For the PRRSV-1 experiments, MA-104 cells were infected with an isolate based on the Porcilis vaccine strain (KJ127878.1 but with several accumulated mutations, see associated publication) and harvested at 8 hpi after pre-treatment with cycloheximide (CHX). For the PRRSV-2 experiments, MARC-145 cells were infected with SD95-21 PRRSV (KC469618.1), and a mutant thereof (KO2). One group of samples was harvested at 9 hpi after pre-treatment with CHX, and another group of samples was harvested at 3, 6, 9 and 12 hpi by flash-freezing without CHX pre-treatment. For all samples, RNA was extracted, ribosomal RNA was removed using Illumina's RiboZero kit, and remaining RNA was gel purified to select fragments 25-34 nt (PRRSV-1-infected samples, CHX-pre-treated PRRSV-2- or mock-infected samples, and non-CHX-pre-treated PRRSV-2- or mock-infected 9 hpi replicate one samples) or ~50 nt long (all other samples). Fragments were cloned into adapters based on the TruSeq small RNA adapters. For all PRRSV-2-infected (or mock-infected) libraries, adapters with an additional seven random nucleotides at the 5′-end of the 3′-adapter and the 3′-end of the 5′-adapter were used. For PRRSV-1 replicate one, no random nucleotides were present on the adapters, and for PRRSV-1 replicate two, 14 random nucleotides were present at the 5′-end of the 3′-adapter. Libraries were sequenced on the Illumina NextSeq 500 platform as a single-end run. For one NextSeq run, some potyvirus amplicons, indexed using TruSeq small RNA indices 1-48, was spiked in to the pool directly before sequencing, therefore some libraries (noCHX_RNA_9hpi_KO2_1, noCHX_RNA_9hpi_KO2_2, noCHX_RNA_9hpi_mock_1, noCHX_RNA_9hpi_mock_2, noCHX_RNA_9hpi_WT_1, noCHX_RNA_9hpi_WT_2) have a small proportion of potyvirus reads, but this does not represent co-infection in the biological sample and does not affect the conclusions of the study. Note that sample nomeclature (including replicate numbers) is consistent between this and the two related accessions, and RiboSeq libraries are matched with RNASeq libraries, which were prepared from the same lysate.
2022-03-09 | E-MTAB-10623 | biostudies-arrayexpress
Project description:Cell lines derived from Chlorocebus sabaeus kidney were infected with an isolate of PRRSV-1 or PRRSV-2 and ribosome profiling was performed (this entry) in parallel with RNASeq (see related accession number). These datasets were used to analyse the viral and host translatome, frameshifting on the viral genome, and putative frameshift-related ribosome pausing events. For the PRRSV-1 experiments, MA-104 cells were infected with an isolate based on the Porcilis vaccine strain (KJ127878.1 but with several accumulated mutations, see associated publication) and harvested at 8 hpi after pre-treatment with cycloheximide (CHX). For the PRRSV-2 experiments, MARC-145 cells were infected with SD95-21 PRRSV (KC469618.1), and a mutant thereof (KO2). One group of samples was harvested at 9 hpi after pre-treatment with CHX, and another group of samples was harvested at 3, 6, 9 and 12 hpi by flash-freezing without CHX pre-treatment. For all samples, RNase I treatment was carried out, following which ribosomes and enclosed RNA were isolated by centrifugation through a sucrose cushion. RNA was extracted, ribosomal RNA was removed using Illumina's RiboZero kit, and remaining RNA was gel purified to select fragments 25-34 nt (PRRSV-1-infected samples, CHX-pre-treated PRRSV-2- or mock-infected samples, and non-CHX-pre-treated PRRSV-2- or mock-infected 9 hpi replicate one samples) or 19-34 nt long (all other samples). Fragments were cloned into adapters based on the TruSeq small RNA adapters. For all PRRSV-2-infected (or mock-infected) libraries, adapters with an additional seven random nucleotides at the 5′-end of the 3′-adapter and the 3′-end of the 5′-adapter were used. For PRRSV-1 replicate one, no random nucleotides were present on the adapters, and for PRRSV-1 replicate two, 14 random nucleotides were present at the 5′-end of the 3′-adapter. Libraries were sequenced on the Illumina NextSeq 500 platform as a single-end run. Non-CHX-pre-treated PRRSV-2-infected 9 hpi replicate two libraries were uploaded under a separate accession number due to differences in the size selection and sequencing protocol - see associated paired-end entry. Note that sample nomeclature (including replicate numbers) is consistent between this and the two related accessions, and RiboSeq libraries are matched with RNASeq libraries, which were prepared from the same lysate. The noCHX_Ribo_9hpi_mock_3 library is deliberately absent as this was a poor quality library.
2022-03-09 | E-MTAB-10621 | biostudies-arrayexpress
Project description:Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV). It clinically manifests as fever, bleeding, and respiratory symptoms in pigs of different age groups, as well as reproductive disorders, causing huge economic losses to the global swine industry. RNA interference (RNAi), a highly conserved mechanism mediating post-transcriptional gene silencing, exerts antiviral effects by cleaving viral double-stranded RNA to generate virus-derived siRNA (vsiRNA). VsiRNA-mediated antiviral immunity is widespread in plants and invertebrates, while in mammals, studies have focused on reducing PRRSV viral load in pigs via siRNA delivery. Therefore, PRRSV strains (LY and LY-R) were primarily selected as virus models to infect PAM and Marc-145 cells, respectively. Then, miRNA and vsiRNA analysis were performed and bioinformatics analysis revealed multiple differentially expressed miRNAs following PRRSV infection. Further enrichment analysis indicated that these miRNAs were mainly enriched in metabolic, endocytic, and phosphorylation signaling pathways. Additionally, in Marc-145 cells infected with PRRSV-LY, high abundance of vsiRNA was detected within the PRRSV-N genome. The N-vsiRNA was inserted into vector psilencer4.1. to construct a PS-N plasmid, which is capable of exerting anti-PRRSV (LY, LY-R, and NADC30-like) activity in Marc-145 cells. Finally, the PS-N plasmid was formulated with PLGA-PEI to prepare a nanoparticle mixture, which was delivered to pigs via intramuscular injection to alleviate tissue damage caused by PRRSV-LY infection to a certain extent. In summary, the porcine RNAi system and vsiRNA generated via viral cleavage play a critical role in inhibiting virus replication. This further demonstrates the feasibility of vsiRNA-based antiviral strategies and offers a reference for the prevention and control of various RNA viruses.
2025-10-01 | GSE308671 | GEO
Project description:MARC-145 cells were infected with porcine reproductive and respiratory syndrome virus (PRRSV)-2 isolate SD95-21 (KC469618.1), and a mutant thereof (KO2), and subjected to ribosome profiling to analyse the viral and host translatome, frameshifting on the viral genome, and putative frameshift-related ribosome pausing events. Samples were harvested at 9 hpi by flash-freezing, without cycloheximide pre-treatment. RNase I treatment was carried out, following which ribosomes and enclosed RNA were isolated by centrifugation through a sucrose cushion. RNA was extracted, ribosomal RNA was removed using Illumina's RiboZero kit, and remaining RNA was gel purified to select fragments 19-80nt long. Fragments were cloned into adapters based on the TruSeq small RNA adapters, with an additional seven random nucleotides at the 5′-end of the 3′-adapter and the 3′-end of the 5′-adapter. Libraries were sequenced on the Illumina NextSeq 500 platform as a paired-end run. Note that sample nomeclature (including replicate numbers) is consistent between this and the two related accessions, and RiboSeq libraries are matched with RNASeq libraries, which were prepared from the same lysate.
2022-03-09 | E-MTAB-10622 | biostudies-arrayexpress