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      Classical swine fever virus replicated poorly in cells from MxA transgenic pigs

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          Abstract

          Background

          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. 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.

          Results

          To 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.

          Conclusions

          Transgene 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.

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          Most cited references22

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          Disruption of the CFTR gene produces a model of cystic fibrosis in newborn pigs.

          Christopher Rogers, David Stoltz, David K. Meyerholz … (2008)
          Almost two decades after CFTR was identified as the gene responsible for cystic fibrosis (CF), we still lack answers to many questions about the pathogenesis of the disease, and it remains incurable. Mice with a disrupted CFTR gene have greatly facilitated CF studies, but the mutant mice do not develop the characteristic manifestations of human CF, including abnormalities of the pancreas, lung, intestine, liver, and other organs. Because pigs share many anatomical and physiological features with humans, we generated pigs with a targeted disruption of both CFTR alleles. Newborn pigs lacking CFTR exhibited defective chloride transport and developed meconium ileus, exocrine pancreatic destruction, and focal biliary cirrhosis, replicating abnormalities seen in newborn humans with CF. The pig model may provide opportunities to address persistent questions about CF pathogenesis and accelerate discovery of strategies for prevention and treatment.
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            High-efficiency thermal asymmetric interlaced PCR for amplification of unknown flanking sequences.

            Yao Liu, Julie Y. Chen (2007)
            Isolation of unknown DNA sequences flanked by known sequences is an important task in molecular biology research. Thermal asymmetric interlaced PCR (TAIL-PCR) is an effective method for this purpose. However the success rate of the original TAIL-PCR needs to be increased, and it is more desirable to obtain target products with larger sizes. Here we present a substantially improved TAIL-PCR procedure with special primer design and optimized thermal conditions. This high-efficiency TAIL-PCR (hiTAIL-PCR) combines the advantages of the TAIL-cycling and suppression-PCR, thus it can block the amplification of nontarget products and suppress small target ones, but allow efficient amplification of large target sequences. Using this method, we isolated genomic flanking sequences of T-DNA insertions from transgenic rice lines. In our tests, the success rates of the reactions were higher than 90%, and in most cases the obtained major products had sizes of 1-3 kb.
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              Conservation of methylation reprogramming in mammalian development: aberrant reprogramming in cloned embryos.

              Diana E. Wolf, Matthew J. Walter, M Stojkovic … (2001)
              Mouse embryos undergo genome-wide methylation reprogramming by demethylation in early preimplantation development, followed by remethylation thereafter. Here we show that genome-wide reprogramming is conserved in several mammalian species and ask whether it also occurs in embryos cloned with the use of highly methylated somatic donor nuclei. Normal bovine, rat, and pig zygotes showed a demethylated paternal genome, suggesting active demethylation. In bovine embryos methylation was further reduced during cleavage up to the eight-cell stage, and this reduction in methylation was followed by de novo methylation by the 16-cell stage. In cloned one-cell embryos there was a reduction in methylation consistent with active demethylation, but no further demethylation occurred subsequently. Instead, de novo methylation and nuclear reorganization of methylation patterns resembling those of differentiated cells occurred precociously in many cloned embryos. Cloned, but not normal, morulae had highly methylated nuclei in all blastomeres that resembled those of the fibroblast donor cells. Our study shows that epigenetic reprogramming occurs aberrantly in most cloned embryos; incomplete reprogramming may contribute to the low efficiency of cloning.
              Article 0 comments Cited 145 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                Yicheng Zhao: yichengzhao@live.cn
                Tiedong Wang: tiedongwang2010@hotmail.com
                Li Yao: yaoli@rnaeditplus.org
                Bo Liu: liubo8510@live.cn
                Chunbo Teng: chunboteng@nefu.edu.cn
                Hongsheng Ouyang: oyhs64@hotmail.com
                Journal
                Journal ID (nlm-ta): BMC Vet Res
                Journal ID (iso-abbrev): BMC Vet. Res
                Title: BMC Veterinary Research
                Publisher: BioMed Central (London )
                ISSN (Electronic): 1746-6148
                Publication date (Electronic): 17 August 2016
                Publication date PMC-release: 17 August 2016
                Publication date Collection: 2016
                Volume: 12
                Electronic Location Identifier: 169
                Affiliations
                [1 ]College of Life Science, Northeast Forestry University, Harbin, China
                [2 ]College of Animal Sciences, Jilin University, Changchun, China
                [3 ]College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China
                Article
                Publisher ID: 794
                DOI: 10.1186/s12917-016-0794-5
                PMC ID: 4987965
                PubMed ID: 27535023
                SO-VID: e58b313b-1a28-4d62-ac1d-a0389a1eb6cb
                Copyright © © Zhao et al. 2016
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                Date received : 26 February 2015
                Date accepted : 9 August 2016
                Categories
                Subject: Methodology Article
                Custom metadata
                issue-copyright-statement © The Author(s) 2016

                ScienceOpen disciplines: Veterinary medicine
                Keywords: classical swine fever virus,mxa,nuclear transplantation,transgenic pig
                Data availability:
                ScienceOpen disciplines: Veterinary medicine
                Keywords: classical swine fever virus, mxa, nuclear transplantation, transgenic pig

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