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Volume 34 Issue 5
October 2019
    Citation: Jian Zhang, Zhenfang Wu, Huaqiang Yang. Aminopeptidase N Knockout Pigs Are Not Resistant to Porcine Epidemic Diarrhea Virus Infection .VIROLOGICA SINICA, 2019, 34(5) : 592-595.  http://dx.doi.org/10.1007/s12250-019-00127-y
    shu

    Aminopeptidase N Knockout Pigs Are Not Resistant to Porcine Epidemic Diarrhea Virus Infection

    • 1.

      National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, China

    • 2.

      Wens Foodstuff Group Co., Ltd, Yunfu 527400, China

    • Corresponding author: Huaqiang Yang, yangh@scau.edu.cn
    • Received Date: 19 December 2018
      Accepted Date: 15 April 2019
      Published Date: 27 May 2019
      Available online: 01 October 2019
    • Porcine epidemic diarrhea virus (PEDV) is the etiologic agent of porcine epidemic diarrhea (PED), which is an acute, highly contagious, and devastating enteric viral disease in pigs. Surface expression of Aminopeptidase N (APN) is considered a functional receptor for infection by porcine epidemic diarrhea virus (PEDV). To test the role of APN in PEDV infection, we created APN knockout (KO) pigs by introducing frame shift mutations in the exon 2 or exon 3 of APN gene using CRIPSR/Cas9-mediated gene editing and somatic cell nuclear transfer. The resulting cloned pigs harbored homozygous or biallelic insertion/deletion in the target site of APN, and had no APN expression in the small intestines. After PEDV infection, both APN KO and wild type controls became infected and showed no differences in clinical signs, fecal viral shedding or antibody response. The results demonstrate that the expression of the APN gene is not necessary for infection of pigs with PEDV and that the absence of APN does not contribute to the resistance to the disease. This work indicates that an alternative receptor is needed to be identified for PEDV infection of pigs.
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      10.1007s12250-019-00127-y-ESM1.pdf

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      2. Lee C (2015) Porcine epidemic diarrhea virus: an emerging and re-emerging epizootic swine virus. Virol J 12:193
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      4. Li W, Li H, Liu Y, Pan Y, Deng F, Song Y, Tang X, He Q (2012) New variants of porcine epidemic diarrhea virus, China, 2011. Emerg Infect Dis 18:1350
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      5. Li W, van Kuppeveld FJM, He Q, Rottier PJM, Bosch BJ (2016) Cellular entry of the porcine epidemic diarrhea virus. Virus Res 226:117-127
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      6. Li W, Luo R, He Q, van Kuppeveld FJM, Rottier PJM, Bosch BJ (2017) Aminopeptidase N is not required for porcine epidemic diarrhea virus cell entry. Virus Res 235:6-13
          doi: 10.1016/j.virusres.2017.03.018

      7. Li W, Hulswit RJG, Kenney SP, Widjaja I, Jung K, Alhamo MA, van Dieren B, van Kuppeveld FJM, Saif LJ, Bosch BJ (2018) Broad receptor engagement of an emerging global coronavirus may potentiate its diverse cross-species transmissibility. Proc Natl Acad Sci U S A 115:E5135-E5143
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      8. Miller LC, Crawford KK, Lager KM, Kellner SG, Brockmeier SL (2016) Evaluation of two real-time polymerase chain reaction assays for Porcine epidemic diarrhea virus (PEDV) to assess PEDV transmission in growing pigs. J Vet Diagn Invest 28:20-29
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      9. Niederwerder MC, Nietfeld JC, Bai J, Peddireddi L, Breazeale B, Anderson J, Kerrigan MA, An B, Oberst RD, Crawford K, Lager KM, Madson DM, Rowland RR, Anderson GA, Hesse RA (2016) Tissue localization, shedding, virus carriage, antibody response, and aerosol transmission of Porcine epidemic diarrhea virus following inoculation of 4-week-old feeder pigs. J Vet Diagn Invest 28:671-678
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      10. Shirato K, Maejima M, Islam MT, Miyazaki A, Kawase M, Matsuyama S, Taguchi F (2016) Porcine aminopeptidase N is not a cellular receptor of porcine epidemic diarrhea virus, but promotes its infectivity via aminopeptidase activity. J Gen Virol 97:2528-2539
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      13. Whitworth KM, Lee K, Benne JA, Beaton BP, Spate LD, Murphy SL, Samuel MS, Mao J, O'Gorman C, Walters EM, Murphy CN, Driver J, Mileham A, McLaren D, Wells KD, Prather RS (2014) Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos. Biol Reprod 91:78

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      15. Whitworth KM, Rowland RRR, Petrovan V, Sheahan M, Cino-Ozuna AG, Fang Y, Hesse R, Mileham A, Samuel MS, Wells KD, Prather RS (2019) Resistance to coronavirus infection in amino peptidase N-deficient pigs. Transgenic Res 28:21-32
          doi: 10.1007/s11248-018-0100-3

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      Aminopeptidase N Knockout Pigs Are Not Resistant to Porcine Epidemic Diarrhea Virus Infection

        Corresponding author: Huaqiang Yang, yangh@scau.edu.cn
      • 1. National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, China
      • 2. Wens Foodstuff Group Co., Ltd, Yunfu 527400, China
      • Received Date: 19 December 2018
      • Accepted Date: 15 April 2019
      • Published Date: 27 May 2019

      Abstract: Porcine epidemic diarrhea virus (PEDV) is the etiologic agent of porcine epidemic diarrhea (PED), which is an acute, highly contagious, and devastating enteric viral disease in pigs. Surface expression of Aminopeptidase N (APN) is considered a functional receptor for infection by porcine epidemic diarrhea virus (PEDV). To test the role of APN in PEDV infection, we created APN knockout (KO) pigs by introducing frame shift mutations in the exon 2 or exon 3 of APN gene using CRIPSR/Cas9-mediated gene editing and somatic cell nuclear transfer. The resulting cloned pigs harbored homozygous or biallelic insertion/deletion in the target site of APN, and had no APN expression in the small intestines. After PEDV infection, both APN KO and wild type controls became infected and showed no differences in clinical signs, fecal viral shedding or antibody response. The results demonstrate that the expression of the APN gene is not necessary for infection of pigs with PEDV and that the absence of APN does not contribute to the resistance to the disease. This work indicates that an alternative receptor is needed to be identified for PEDV infection of pigs.

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