A possible strategy to produce pigs resistant to porcine reproductive and respiratory syndrome virus

Luo, Biping; Ju, Shiqiang; Wang, Bin; Ren, Rui

doi:10.1016/j.antiviral.2013.05.010

Cited by 3 publications

(3 citation statements)

References 46 publications

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“…Specific gene silencing also can be triggered in mammalian cells by using synthetic short interfering RNA (siRNA), and plasmid or virus-mediated short hairpin RNA (shRNA). These RNAi strategies have been used successfully for suppressing the replication of human and animal viruses, including hepatitis B virus (HBV) (Kayhan et al, 2007), human immunodeficiency virus type (HIV) (Coburn and Cullen, 2002), porcine reproductive and respiratory syndrome virus (PRRSV) (Luo et al, 2013) and foot-and-mouth disease virus (FMDV) (De los Santos et al, 2005;Lv et al, 2009;Xu et al, 2012). Furthermore, we and others reported that transgenic animals expressing shRNA against virus genes showed a significant resistance to viral challenge (Pengyan et al, 2010;Li et al, 2014;Daniel-Carlier et al, 2013;Du et al, 2014).…”

Section: Introductionmentioning

confidence: 94%

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Qiao

et al. 2015

Journal of Virological Methods

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Section: Introductionmentioning

confidence: 94%

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Qiao

et al. 2015

Journal of Virological Methods

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“…After culture for 7-10 days, G418-resistant cell colonies expressing EGFP, as assessed by fluorescence microscopy, were isolated separately. Each single colony was seeded separately in cell culture medium with G418 (300 lg/mL) to expand the stable integrated cell population, and then cryopreserved in liquid nitrogen for further use in SCNT (Luo et al, 2013).…”

Section: Preparation Of Nuclear Donor Cellsmentioning

confidence: 99%

“…SCNT procedures were conducted as previously described ( Ju et al, 2010;Luo et al, 2013). Briefly, all manipulations were performed in microdrops of HEPES-buffered TCM-199 medium supplemented with 0.3% bovine serum albumin (BSA) and 7.5 lg/mL cytochalasin B covered with mineral oil.…”

Section: Nuclear Transfer and Culture Of Scnt Embryosmentioning

confidence: 99%

Effects of Histone Acetylation Status on the Early Development ofIn VitroPorcine Transgenic Cloned Embryos

Luo

Muneri

et al. 2015

Cellular Reprogramming

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The purpose of this study was to investigate the effects of the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) on transgene expression and development of porcine transgenic cloned embryos, specifically focusing on effects derived from TSA-treated donor cells or TSA-treated reconstructed embryos. The results showed that TSA treatment on reconstructed embryos modified the acetylation status, which significantly improved the development of porcine somatic cell nuclear transfer (SCNT) embryos in vitro, but not donor cells. Furthermore, the treatment of reconstructed embryos with TSA enhanced expression of the pluripotency-related gene POU5F1 and stimulated expression of the anti-apoptotic gene BCL-2. Enhanced green fluorescent protein (EGFP) mRNA expression of every group dropped drastically from donor cells to blastocysts. Interestingly, TSA is likely to prevent a decline in EGFP expression in nuclear reprogramming of porcine SCNT embryos. However DNA hypomethylation induced by modified histone acetylation of donor cells treated with TSA was significantly more effective in increasing EGFP expression in SCNT blastocysts. In conclusion, the acetylation status of both donor cells and reconstructed embryos modified by TSA treatment increased transgene expression and improved nuclear reprogramming and the developmental potential of porcine transgenic SCNT embryos.

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Inhibition of porcine reproductive and respiratory syndrome virus replication in vitro using DNA-based short antisense oligonucleotides

Zheng

Li²,

Zhu

et al. 2015

BMC Vet Res

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BackgroundPorcine reproductive and respiratory syndrome (PRRS) is caused by porcine reproductive and respiratory syndrome virus (PRRSV) and is an economically important disease in swine-producing areas. The objective of this study was to screen for effective antisense oligonucleotides (AS-ONs) which could inhibit PRRSV replication in MARC-145 cells and in pulmonary alveolar macrophages (PAM).ResultsNine short AS-ON sequences against the well-conserved regions of PRRSV (5′-UTR, NSP9, ORF5 and ORF7) were selected. When MARC-145 cells or PAM were infected with PRRSV followed by transfection with AS-ONs, four AS-ON sequences targeting 5′-UTR, ORF5 or NSP9 were found to be the most effective oligonucleotides in decreasing the cytopathic effect (CPE) induced by PRRSV infection. Quantitative PCR and indirect immunofluorescence staining confirmed that ORF7 levels were significantly reduced both at RNA and protein levels. The PRRSV titration data furthermore indicated that transfection with AS-ON YN8 could reduce the PRRSV titer by 1000-fold compared with controls.ConclusionThe results presented here indicate that DNA-based antisense oligonucleotides can effectively inhibit PRRSV replication in MARC-145 cells and in PAM. Furthermore, comparing with the reported hit rates (approximately 10-30 %), we achieved a higher success rate (44 %). The strategy we took to design the antisense sequences might be applied to select AS-ONs that more efficiently reduce the expression of target genes.

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A possible strategy to produce pigs resistant to porcine reproductive and respiratory syndrome virus

Cited by 3 publications

References 46 publications

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference

Effects of Histone Acetylation Status on the Early Development ofIn VitroPorcine Transgenic Cloned Embryos

Inhibition of porcine reproductive and respiratory syndrome virus replication in vitro using DNA-based short antisense oligonucleotides

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