Abstract
BackgroundIn addition to their value as livestock, pigs are susceptible to classical swine fever virus (CSFV) and can serve as reservoirs for CSFV, allowing it to develop into an epizootic. CSFV, a pestivirus of the Flaviviridae family, has a single-stranded RNA genome. Recent research has indicated that the human MxA protein inhibits the life cycles of certain RNA viruses, such as members of the Bunyaviridae family, the Flaviviridae family and others.ResultsTo produce pigs with antiviral protection against CSFV, transgenic pigs expressing human MxA were generated by nuclear transplantation. Cells from three MxA transgenic piglets were used to investigate in vitro antiviral activity of MxA aganist CSFV, and the results of in vitro indirect immunofluorescence assays, virus titration and real-time PCR indicated that the MxA transgenic pig has an antiviral capacity against CSFV.ConclusionsTransgene with human MxA on pigs is feasible. High levels of MxA expression do inhibit CSFV in vitro at early time points post-infection at 60-96dpi.
Highlights
In addition to their value as livestock, pigs are susceptible to classical swine fever virus (CSFV) and can serve as reservoirs for CSFV, allowing it to develop into an epizootic
The No 34 clone (Fig. 1a) was chosen for Nuclear transplantation (NT), about 500 SCNT embryos were transferred into 3 surrogate mothers that exhibited natural estrus and three male transgenic piglets were obtained by natural delivery on day 119 post-implantation (Fig. 1b) and one piglet died before birth, the other three live transgenic piglets displayed no obvious weight differences comparing to a normal male piglet born at the same day (Fig. 1c)
The sequence analysis revealed that the MxA fragment had randomly inserted into introns on chromosome 16 (, NW_003612429:30482-30588)
Summary
In addition to their value as livestock, pigs are susceptible to classical swine fever virus (CSFV) and can serve as reservoirs for CSFV, allowing it to develop into an epizootic. CSFV, a pestivirus of the Flaviviridae family, has a single-stranded RNA genome. Human MxA has been shown to exhibit a wide antiviral capacity against small RNA viruses. This interferoninduced protein is one of the best-studied determinants of innate immunity to viral infection [1]. MxA belongs to the dynamin superfamily of large GTPases and has a C-terminal stretch of basic amino acids that constitutes a nuclear localization signal (NLS) This NLS mediates the nuclear accumulation of MxA, whereas the swine Mx1 protein is not located inside nucleus for lack of NLS. Classical swine fever virus (CSFV) is a pestivirus of the Flaviviridae family [8] with an enveloped virion incorporating glycosylated membrane proteins. The CSFV genome has a single strand of positive-sense RNA [9]
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