Syncrip facilitates porcine parvovirus viral dna replication through the alternative splicing of ns1 mrna to promote ns2 mrna formation

Document Outline

• SYNCRIP facilitates porcine parvovirus viral DNA replication through the alternative splicing of NS1 mRNA to promote NS2 mRNA formation
  • Abstract 
  • Introduction
  • Materials and methods
    • Cell culture and virus
    • Plasmid constructions and reactions
    • Antibodies and reagents
    • CRISPR-Cas9 mediated SYNCRIP knockout in PK-15 cells
    • Reverse transcription quantitative PCR
    • Immunofluorescence assay and confocal imaging
    • RNA-immunoprecipitation assay
    • RNA-pulldown assay
    • Electrophoretic mobility shift assay
    • Purification of GST-tagged SYNCRIP protein
    • Statistics
  • Results
    • PPV NS1 mRNA can specifically interact with SYNCRIP
    • PPV infection up-regulates SYNCRIP expression in vitro and in vivo
    • CRISPR-Cas9 mediated knockout of SYNCRIP impairs PPV replication
    • Over-expression of SYNCRIP leads to the reduction of NS1 mRNA and protein expression
    • SYNCRIP regulates the ratio of NS1 mRNA to NS2 mRNA and the replication of PPV
    • SYNCRIP affects the NS1 expression by acting on the 3′-terminal sites of NS2 mRNA
  • Discussion
  • Acknowledgements
  • References