Project description:In this study, we generated the PBMC transcriptome data of 3 Landrace, and 3 indigenous Ghurrah pigs before and 7 days after Classical Swine fever vaccination. The piglets were 8-12 weeks of age. The sequencing was done on Illumina X-Ten platform, generating average of 39 million 150 bp paired end reads for each sample.
Project description:African swine fever virus is highly contagious and causes a fatal infectious disease in pigs, resulting in a significant global impact on pork supply. The African swine fever virus RNA polymerase serves as a crucial multifunctional protein complex responsible for genome transcription and regulation. Therefore, it is essential to investigate its structural and functional characteristics for the prevention and control of African swine fever. Here, we determine the structures of endogenous African swine fever virus RNA polymerase in both nucleic acid-free and elongation states. The African swine fever virus RNA polymerase shares similarities with the core of typical RNA polymerases, but possesses a distinct subunit M1249L. Notably, the dynamic binding mode of M1249L with RNA polymerase, along with the C-terminal tail insertion of M1249L in the active center of DNA-RNA scaffold binding, suggests the potential of M1249L to regulate RNA polymerase activity within cells. These results are important for understanding the transcription cycle of African swine fever virus and for developing antiviral strategies.
Project description:Classical swine fever virus (CSFV) is an etiologic agent that causes a highly contagious disease in pigs. Laying a foundation to solve problems in its pathogenic mechanism, microarray analysis was performed to detect the gene transcriptional profiles in peripheral blood mononuclear cells (PBMC) following infection with a Chinese highly virulent CSFV strain Shimen. Three susceptible pigs were inoculated intramuscularly with a lethal dose (1.0 × 106 TCID50) of CSFV. Pigs showed classical CSF signs, depletion of lymphocytes and monocytes consistent with CSFV infection, and the CSFV genome was also confirmed in the PBMC. The PBMC were isolated at 1, 3, 6 and 9 days post-inoculation (dpi). Total RNA were extracted and subjected to microarray analysis. Data showed that expression of 847 genes wherein 467 genes were known function and the remaining 380 genes were unknown function, and 541 up- and 306 down-regulation, altered after infection. There were 54, 181, 438 and 354 up- and 61, 120, 218 and 145 down-regulated genes presented on 1, 3, 6 and 9 dpi, respectively. These genes were involved in immune response (14.5%), apoptosis (3.3%), signal transduction (7.6%), transcription (4.4%), metabolism (11%), transport (3.9%), development (6.8%) and cell cycle (3.7%). Results demonstrated its usefulness in exploring the pathogenic mechanisms of CSFV.
Project description:African swine fever virus (ASFV) is a lethal animal pathogen which enters its host cells through endocytosis. So far, host factors specifically required for ASFV replication have been barely identified. In this study a genome-wide CRISPR/Cas9 knockout screen in porcine cells indicated that the genes RFXANK, RFXAP, SLA-DMA, SLA-DMB, and CIITA are important for productive ASFV infection. The proteins encoded by these genes belong to the major-histocompatibility-complex II (MHC II), or swine-leucocyte-antigen-complex II (SLA II). RFXAP and CIITA are MHC II-specific transcription factors, whereas SLA-DMA/B are subunits of the non-classical MHC II molecule SLA-DM. Targeted knockout of either of these genes led to severe replication defects of different ASFV isolates, reflected by substantially reduced plating efficiency, cell-to-cell spread, and progeny virus titers. For the characterization of the knockouts on a proteome level the protein contents of the knockout cell lines were analyzed by mass spectrometry.