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Transgenic shRNA pigs reduce susceptibility to foot and mouth disease virus infection.

Hu S, Qiao J, Fu Q, Chen C, Ni W, Wujiafu S, Ma S, Zhang H, Sheng J, Wang P, Wang D, Huang J, Cao L, Ouyang H - Elife (2015)

Bottom Line: Foot-and-mouth disease virus (FMDV) is an economically devastating viral disease leading to a substantial loss to the swine industry worldwide.In vitro challenge of TG fibroblasts showed the shRNA suppressed viral growth.Our results show that TG shRNA can provide a viable tool for producing animals with enhanced resistance to FMDV.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Shihezi University, Shihezi, China.

ABSTRACT
Foot-and-mouth disease virus (FMDV) is an economically devastating viral disease leading to a substantial loss to the swine industry worldwide. A novel alternative strategy is to develop pigs that are genetically resistant to infection. Here, we produce transgenic (TG) pigs that constitutively expressed FMDV-specific short interfering RNA (siRNA) derived from small hairpin RNA (shRNA). In vitro challenge of TG fibroblasts showed the shRNA suppressed viral growth. TG and non-TG pigs were challenged by intramuscular injection with 100 LD50 of FMDV. High fever, severe clinical signs of foot-and-mouth disease and typical histopathological changes were observed in all of the non-TG pigs but in none of the high-siRNA pigs. Our results show that TG shRNA can provide a viable tool for producing animals with enhanced resistance to FMDV.

No MeSH data available.


Related in: MedlinePlus

Body temperature curve of all infected pigs.The body temperatures of non-TG (120, 141, 159, 191 and 211) and TG pigs (11, 19, 24, 49 and 78) were measured at 24 hr intervals before FMDV challenge until the end of the experiment.DOI:http://dx.doi.org/10.7554/eLife.06951.008
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fig4s1: Body temperature curve of all infected pigs.The body temperatures of non-TG (120, 141, 159, 191 and 211) and TG pigs (11, 19, 24, 49 and 78) were measured at 24 hr intervals before FMDV challenge until the end of the experiment.DOI:http://dx.doi.org/10.7554/eLife.06951.008

Mentions: The resistance of TG pigs to FMDV infection was further tested by intramuscular injection of O serotypes of FMDV. The challenged animals included high-siRNA TG (11 and 19), low-siRNA TG (24, 49 and 78) and non-TG pigs (n=5). Prior to the day of infection, no animal tested was positive for FMDV. After FMDV challenge, all non-TG pigs developed high fever within 72 hr of challenge and severe clinical signs of FMD, the appearance of vesicles on the feet and nose (Figure 4A and Figure 4—figure supplement 1). All non-TG pigs became deteriorated and the lesion score reached 24 at 5 d after challenge (Figure 4A and Figure 4—figure supplement 1). Some smaller vesicles in low-siRNA TG (24, 49 and 78) pigs were also observed until 7 d after challenge, as shown in Figure 4A. However, the body temperature of high-siRNA TG (11 and 19) pigs remained normal throughout the experiment (Figure 4A and Figure 4—figure supplement 1). TG pigs 11 and 19 developed one small vesicle at day 9 of challenge, but TG pig 11 recovered soon on the next day (Figure 4A). We subsequently quantified the viral genome RNA in the serum of the infected animals. Consistent with the clinical signs data, the viral load in the serum of the high-siRNA TG and low-siRNA TG pigs was lower than that in the non-TG pigs (Figure 4B). The viral RNA expression in serum was 42-fold lower in the high-siRNA TG group than that in the non-TG pigs at day 10 of challenge.10.7554/eLife.06951.007Figure 4.Transgenic shRNA pigs resisted FMDV infection.


Transgenic shRNA pigs reduce susceptibility to foot and mouth disease virus infection.

Hu S, Qiao J, Fu Q, Chen C, Ni W, Wujiafu S, Ma S, Zhang H, Sheng J, Wang P, Wang D, Huang J, Cao L, Ouyang H - Elife (2015)

Body temperature curve of all infected pigs.The body temperatures of non-TG (120, 141, 159, 191 and 211) and TG pigs (11, 19, 24, 49 and 78) were measured at 24 hr intervals before FMDV challenge until the end of the experiment.DOI:http://dx.doi.org/10.7554/eLife.06951.008
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4502569&req=5

fig4s1: Body temperature curve of all infected pigs.The body temperatures of non-TG (120, 141, 159, 191 and 211) and TG pigs (11, 19, 24, 49 and 78) were measured at 24 hr intervals before FMDV challenge until the end of the experiment.DOI:http://dx.doi.org/10.7554/eLife.06951.008
Mentions: The resistance of TG pigs to FMDV infection was further tested by intramuscular injection of O serotypes of FMDV. The challenged animals included high-siRNA TG (11 and 19), low-siRNA TG (24, 49 and 78) and non-TG pigs (n=5). Prior to the day of infection, no animal tested was positive for FMDV. After FMDV challenge, all non-TG pigs developed high fever within 72 hr of challenge and severe clinical signs of FMD, the appearance of vesicles on the feet and nose (Figure 4A and Figure 4—figure supplement 1). All non-TG pigs became deteriorated and the lesion score reached 24 at 5 d after challenge (Figure 4A and Figure 4—figure supplement 1). Some smaller vesicles in low-siRNA TG (24, 49 and 78) pigs were also observed until 7 d after challenge, as shown in Figure 4A. However, the body temperature of high-siRNA TG (11 and 19) pigs remained normal throughout the experiment (Figure 4A and Figure 4—figure supplement 1). TG pigs 11 and 19 developed one small vesicle at day 9 of challenge, but TG pig 11 recovered soon on the next day (Figure 4A). We subsequently quantified the viral genome RNA in the serum of the infected animals. Consistent with the clinical signs data, the viral load in the serum of the high-siRNA TG and low-siRNA TG pigs was lower than that in the non-TG pigs (Figure 4B). The viral RNA expression in serum was 42-fold lower in the high-siRNA TG group than that in the non-TG pigs at day 10 of challenge.10.7554/eLife.06951.007Figure 4.Transgenic shRNA pigs resisted FMDV infection.

Bottom Line: Foot-and-mouth disease virus (FMDV) is an economically devastating viral disease leading to a substantial loss to the swine industry worldwide.In vitro challenge of TG fibroblasts showed the shRNA suppressed viral growth.Our results show that TG shRNA can provide a viable tool for producing animals with enhanced resistance to FMDV.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Shihezi University, Shihezi, China.

ABSTRACT
Foot-and-mouth disease virus (FMDV) is an economically devastating viral disease leading to a substantial loss to the swine industry worldwide. A novel alternative strategy is to develop pigs that are genetically resistant to infection. Here, we produce transgenic (TG) pigs that constitutively expressed FMDV-specific short interfering RNA (siRNA) derived from small hairpin RNA (shRNA). In vitro challenge of TG fibroblasts showed the shRNA suppressed viral growth. TG and non-TG pigs were challenged by intramuscular injection with 100 LD50 of FMDV. High fever, severe clinical signs of foot-and-mouth disease and typical histopathological changes were observed in all of the non-TG pigs but in none of the high-siRNA pigs. Our results show that TG shRNA can provide a viable tool for producing animals with enhanced resistance to FMDV.

No MeSH data available.


Related in: MedlinePlus