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siRNAs targeting PB2 and NP genes potentially inhibit replication of Highly Pathogenic H5N1 Avian Influenza Virus

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Abstract

Highly Pathogenic Avian Influenza (HPAI) H5N1 virus is a threat to animal and public health worldwide. Till date, the H5N1 virus has claimed 402 human lives, with a mortality rate of 58% and has caused the death or culling of millions of poultry since 2003. In this study, we have designed three siRNAs (PB2-2235, PB2-479 and NP-865) targeting PB2 and NP genes of avian influenza virus and evaluated their potential, measured by hemagglutination (HA), plaque reduction and Real time RT-PCR assay, in inhibiting H5N1 virus (A/chicken/Navapur/7972/2006) replication in MDCK cells. The siRNAs caused 8- to 16-fold reduction in virus HA titers at 24 h after challenged with 100TCID50 of virus. Among these siRNAs, PB2-2235 offered the highest inhibition of virus replication with 16-fold reduction in virus HA titer, 80% reduction in viral plaque counts and 94% inhibition in expression of specific RNA at 24 h. The other two siRNAs had 68–73% and 87–88% reduction in viral plaque counts and RNA copy number, respectively. The effect of siRNA on H5N1 virus replication continued till 48h (maximum observation period). These findings suggest that PB2-2235 could efficiently inhibit HPAI H5N1 virus replication.

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Acknowledgements

We thank the Director, Indian Veterinary Research Institute and Indian Council of Agricultural Research, New Delhi, for providing necessary facilities to carry out this work. We are thankful to the Department of Animal Husbandry, Dairying and Fisheries, Ministry of Agriculture, India, for financial support through CDDL for Avian influenza diagnosis and research grant.

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Authors and Affiliations

  1. ICAR-National Institute of High Security Animal Diseases, Bhopal, 462 022, India

    Padmanava Behera, Shanmugasundaram Nagarajan, Harshad V Murugkar, Semmannan Kalaiyarasu, Shiv Chandra Dubey, Diwakar D Kulkarni & Chakradhar Tosh

  2. Department of Microbiology, Barkatullah University, Bhopal, 462 026, India

    Anil Prakash

  3. Department of Biotechnology, Barkatullah University, Bhopal, 462 026, India

    Ragini Gothalwal

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  1. Padmanava Behera
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  2. Shanmugasundaram Nagarajan
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  3. Harshad V Murugkar
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  4. Semmannan Kalaiyarasu
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  5. Anil Prakash
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  6. Ragini Gothalwal
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  7. Shiv Chandra Dubey
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  8. Diwakar D Kulkarni
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  9. Chakradhar Tosh
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Correspondence to Chakradhar Tosh.

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Corresponding editor: Indranil Dasgupta

[Behera P, Nagarajan S, Murugkar HV, Kalaiyarasu S, Prakash A, Gothalwal R, Dubey SC, Kulkarni DD and Tosh C 2015 siRNAs targeting PB2 and NP genes potentially inhibit replication of Highly Pathogenic H5N1 Avian Influenza Virus. J. Biosci. 40 1–8] DOI 10.1007/s12038-015-9524-6

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Behera, P., Nagarajan, S., Murugkar, H.V. et al. siRNAs targeting PB2 and NP genes potentially inhibit replication of Highly Pathogenic H5N1 Avian Influenza Virus . J Biosci 40, 233–240 (2015). https://doi.org/10.1007/s12038-015-9524-6

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