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Production of transgenic pigs over-expressing the antiviral gene Mx1.

Yan Q, Yang H, Yang D, Zhao B, Ouyang Z, Liu Z, Fan N, Ouyang H, Gu W, Lai L - Cell Regen (Lond) (2014)

Bottom Line: It is therefore an interesting candidate gene to improve disease resistance in farm animals.Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells.Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

ABSTRACT
The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interesting candidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs express approximately 15-25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was also markedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs with influenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found that the Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These results demonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate that the Mx1 transgene can be harnessed to develop disease-resistant pigs.

No MeSH data available.


Related in: MedlinePlus

Protective effect of Mx1 transgene against CSFV infection. (A) IFA and FITC conjugated antibodies were used to examine the viral infection in fibroblasts from 5 transgenic and an age-matched non-transgenic pigs; 4 × magnification. (B) Quantification of CSFV-positive cells. Values represent the mean ± s.d., n = 3. *P < 0.01 vs non-transgenic.
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Fig6: Protective effect of Mx1 transgene against CSFV infection. (A) IFA and FITC conjugated antibodies were used to examine the viral infection in fibroblasts from 5 transgenic and an age-matched non-transgenic pigs; 4 × magnification. (B) Quantification of CSFV-positive cells. Values represent the mean ± s.d., n = 3. *P < 0.01 vs non-transgenic.

Mentions: Finally we wanted to study the inhibitory effects of the Mx1 transgene on a virus other than influenza. To this end, we infected fibroblast cells with CSFV and examined its replication by detecting levels of the virus by IFA using anti-CSFV serum. At 60 hours post-CSFV infection, the fibroblasts from non-transgenic controls exhibited bright green fluorescence in the cytoplasm, indicating that most cells were producing the virus (Figure 6). By contrast, fewer cells from Mx1 transgenic pigs displayed green fluorescence. We observed some variation between cells derived from different pigs. Cells from pigs #4-1, #4-2, #4-4 displayed the lowest degree of green fluorescence, indicating that the inhibitory effects on CSFV replication is strongest in these cells (Figure 6).Figure 6


Production of transgenic pigs over-expressing the antiviral gene Mx1.

Yan Q, Yang H, Yang D, Zhao B, Ouyang Z, Liu Z, Fan N, Ouyang H, Gu W, Lai L - Cell Regen (Lond) (2014)

Protective effect of Mx1 transgene against CSFV infection. (A) IFA and FITC conjugated antibodies were used to examine the viral infection in fibroblasts from 5 transgenic and an age-matched non-transgenic pigs; 4 × magnification. (B) Quantification of CSFV-positive cells. Values represent the mean ± s.d., n = 3. *P < 0.01 vs non-transgenic.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4230515&req=5

Fig6: Protective effect of Mx1 transgene against CSFV infection. (A) IFA and FITC conjugated antibodies were used to examine the viral infection in fibroblasts from 5 transgenic and an age-matched non-transgenic pigs; 4 × magnification. (B) Quantification of CSFV-positive cells. Values represent the mean ± s.d., n = 3. *P < 0.01 vs non-transgenic.
Mentions: Finally we wanted to study the inhibitory effects of the Mx1 transgene on a virus other than influenza. To this end, we infected fibroblast cells with CSFV and examined its replication by detecting levels of the virus by IFA using anti-CSFV serum. At 60 hours post-CSFV infection, the fibroblasts from non-transgenic controls exhibited bright green fluorescence in the cytoplasm, indicating that most cells were producing the virus (Figure 6). By contrast, fewer cells from Mx1 transgenic pigs displayed green fluorescence. We observed some variation between cells derived from different pigs. Cells from pigs #4-1, #4-2, #4-4 displayed the lowest degree of green fluorescence, indicating that the inhibitory effects on CSFV replication is strongest in these cells (Figure 6).Figure 6

Bottom Line: It is therefore an interesting candidate gene to improve disease resistance in farm animals.Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells.Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls.

View Article: PubMed Central - PubMed

Affiliation: Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

ABSTRACT
The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interesting candidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cell nuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs express approximately 15-25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was also markedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs with influenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profound decrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-fold reduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found that the Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These results demonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate that the Mx1 transgene can be harnessed to develop disease-resistant pigs.

No MeSH data available.


Related in: MedlinePlus