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The plasmid constructs producing shRNA corresponding to the conserved 3D polymerase of Foot and Mouth Disease virus protects guinea pigs against challenge virus

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Abstract

RNA interference (RNAi) has been used as an effective antiviral strategy for its specific silencing of viral gene expression in mammalian cells. In this study, shRNA targeting two regions of Foot and Mouth Disease Virus (FMDV) i.e. 3D and 5′UTR which are very essential in virus replication were evaluated. The constructs were made using h7K RNA polymerase III promoter. We investigated in vivo inhibitory effect of shRNA on FMDV replication in BHK-21 cells and guinea pigs. The results showed that transfection of 3D shRNA could reduce virus growth by three folds when cells were challenged with 102 TCID50 of FMDV. Pretreated guinea pigs with 3DshRNA were protected 80% with 103 GPID50 of FMDV. As a first report in guinea pigs which are recognized animal model for FMD vaccine potency testing, the study suggests that shRNA could be a viable therapeutic approach to control severity of FMD infection and spread.

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Abbreviations

FMDV:

Foot and Mouth Disease

RNAi:

RNA interference

shRNA:

small hair pin RNA

UTR:

untranslated region

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

cDNA:

complementary DNA

CPE:

cytopathic effect

E.Coli :

Escherichia coli

TCID:

Tissue culture infective dose

GPID:

Guinea pig infective dose

PCR:

polymerase chain reaction

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Acknowledgement

The authors are very much grateful to the Director, Indian Veterinary Research Institute (IVRI), Bareilly and the Joint Director, Bangalore Campus, India for providing the facilities to carry out this research work. The work has been supported and funded by Department of Biotechnology, Govt.of India.

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Authors and Affiliations

  1. Indian Veterinary Research Institute, Molecular Virology Lab, Bangalore, campus, Hebbal, Bangalore, 560024, India

    Dechamma Hosur Joyappa, Sarika Sasi, Kumar C. Ashok, Golla Rama Reddy & Veluvarthy V. S. Suryanarayana

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  1. Dechamma Hosur Joyappa
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  2. Sarika Sasi
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  3. Kumar C. Ashok
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  4. Golla Rama Reddy
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  5. Veluvarthy V. S. Suryanarayana
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Correspondence to Veluvarthy V. S. Suryanarayana.

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Joyappa, D.H., Sasi, S., Ashok, K.C. et al. The plasmid constructs producing shRNA corresponding to the conserved 3D polymerase of Foot and Mouth Disease virus protects guinea pigs against challenge virus. Vet Res Commun 33, 263–271 (2009). https://doi.org/10.1007/s11259-008-9174-3

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  • DOI: https://doi.org/10.1007/s11259-008-9174-3

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