Browse > Article
http://dx.doi.org/10.4142/jvs.21174

Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus  

Hartawan, Risza (Biomedical Science, Faculty of Medicine, Universitas Indonesia)
Pujianto, Dwi Ari (Department of Medical Biology Pre Clinic, Faculty of Medicine, Universitas Indonesia)
Dharmayanti, Ni Luh Putu Indi (Indonesian Research Center for Veterinary Science, Ministry of Agriculture)
Soebandrio, Amin (Eijkman Institute, Ministry of Research, Technology and Higher Education)
Publication Information
Journal of Veterinary Science / v.23, no.2, 2022 , pp. 24.1-24.10 More about this Journal
Abstract
Background: Small interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia. Objectives: The main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus. Methods: The effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells. Results: The siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene. Conclusions: These findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.
Keywords
influenza in birds; H5N1 subtype; Indonesia; RNA interference; antiviral agents;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Spackman E, Killian ML. Avian influenza virus isolation, propagation, and titration in embryonated chicken eggs. In: Spackman E, editor. Animal Influenza Virus. 2nd ed. New York: Humana Press; 2014, 125-140.
2 Whittaker GR. Intracellular trafficking of influenza virus: clinical implications for molecular medicine. Expert Rev Mol Med. 2001;2001(5):1-13.   DOI
3 Earhart KC, Elsayed NM, Saad MD, Gubareva LV, Nayel A, Deyde VM, Abdelsattar A, Abdelghani AS, Boynton BR, Mansour MM, Essmat HM, Klimov A, Shuck-Lee D, Monteville MR, Tjaden JA. Oseltamivir resistance mutation N294S in human influenza A(H5N1) virus in Egypt. J Infect Public Health. 2009;2(2):74-80.   DOI
4 Ge Q, McManus MT, Nguyen T, Shen CH, Sharp PA, Eisen HN, Chen J. RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc Natl Acad Sci U S A. 2003;100(5):2718-2723.   DOI
5 de Jong MD, Tran TT, Truong HK, Vo MH, Smith GJ, Nguyen VC, Bach VC, Phan TQ, Do QH, Guan Y, Peiris JS, Tran TH, Farrar J. Oseltamivir resistance during treatment of influenza A (H5N1) infection. N Engl J Med. 2005;353(25):2667-2672.   DOI
6 Gavrilov K, Saltzman WM. Therapeutic siRNA: principles, challenges, and strategies. Yale J Biol Med. 2012;85(2):187-200.
7 Dharmayanti NLPI, Hartawan R, PudjiatmokoWibawa H, HardimanBalish A, Donis R, Davis CT, Samaan G. Genetic characterization of clade 2.3.2.1 avian influenza A(H5N1) viruses, Indonesia, 2012. Emerg Infect Dis. 2014;20(4):671-674.   DOI
8 Dharmayanti NLPI, Hartawan R, Hewajuli DA. Development of primers for reverse transcriptase polymerase chain reaction to detect avian influenza virus in Indonesia. J Vet. 2016;17:183-196.
9 Suarez DL. Influenza A virus, In: Swayne DE, editor. Animal Influenza. 2nd ed. Iowa: John Wiley & Sons, Inc.; 2016, 3-30.
10 Dharmayanti NLPI, Thor SW, Zanders N, Hartawan R, Ratnawati A, Jang Y, Rodriguez M, Suarez DL, Samaan G, Pudjiatmoko , Davis CT. Attenuation of highly pathogenic avian influenza A(H5N1) viruses in Indonesia following the reassortment and acquisition of genes from low pathogenicity avian influenza A virus progenitors. Emerg Microbes Infect. 2018;7(1):147.   DOI
11 Pawitan JA. Molecular pathogenesis of avian influenza and prospect of theraphy using small interfering RNA. In: Haugan S, Bjornson W, editors. Avian Influenza: Etiology, Pathogenesis and Interventions. New York: Nova Science Publishing; 2010, 69-82.
12 Linke LM, Wilusz J, Pabilonia KL, Fruehauf J, Magnuson R, Olea-Popelka F, Triantis J, Landolt G, Salman M. Inhibiting avian influenza virus shedding using a novel RNAi antiviral vector technology: proof of concept in an avian cell model. AMB Express. 2016;6(1):16.   DOI
13 Sims LD, Brown IH. Multi-continental panzootic of H5 highly pathogenic avian influenza (1996-2015). In: Swayne DE, editor. Animal Influenza. 2nd ed. Iowa: John Wiley & Sons, Inc.; 2016, 204-247.
14 Dharmayanti NLPI, Hewajuli DA, Ratnawati A, Hartawan R. Genetic diversity of the H5N1 viruses in live bird markets, Indonesia. J Vet Sci. 2020;21(4):e56.   DOI
15 Shaw ML, Palese P. Orthomyxoviridae. In: Knipe DM, Howley PM, editors. Fields Virology. 6th ed. Baltimore: Wolter Luwer; 2013, 1151-1185.
16 Gasparini R, Amicizia D, Lai PL, Bragazzi NL, Panatto D. Compounds with anti-influenza activity: present and future of strategies for the optimal treatment and management of influenza. Part II: future compounds against influenza virus. J Prev Med Hyg. 2014;55(4):109-129.
17 Saladino R, Barontini M, Crucianelli M, Nencioni L, Sgarbanti R, Palamara AT. Current advances in anti-influenza therapy. Curr Med Chem. 2010;17(20):2101-2140.   DOI
18 Okomo-Adhiambo M, Fry AM, Su S, Nguyen HT, Elal AA, Negron E, Hand J, Garten RJ, Barnes J, Xiyan X, Villanueva JM, Gubareva LV2013-14 US Influenza Antiviral Working Group. Oseltamivir-resistant influenza A(H1N1)pdm09 viruses, United States, 2013-14. Emerg Infect Dis. 2015;21(1):136-141.   DOI
19 Behera P, Nagarajan S, Murugkar HV, Kalaiyarasu S, Prakash A, Gothalwal R, Dubey SC, Kulkarni DD, Tosh C. siRNAs targeting PB2 and NP genes potentially inhibit replication of highly pathogenic H5N1 avian influenza virus. J Biosci. 2015;40(2):233-240.   DOI
20 Khantasup K, Kopermsub P, Chaichoun K, Dharakul T. Targeted small interfering RNAimmunoliposomes as a promising therapeutic agent against highly pathogenic avian influenza A (H5N1) virus infection. Antimicrob Agents Chemother. 2014;58(5):2816-2824.   DOI
21 Tompkins SM, Lo CY, Tumpey TM, Epstein SL. Protection against lethal influenza virus challenge by RNA interference in vivo. Proc Natl Acad Sci U S A. 2004;101(23):8682-8686.   DOI
22 He G, Qiao J, Dong C, He C, Zhao L, Tian Y. Amantadine-resistance among H5N1 avian influenza viruses isolated in Northern China. Antiviral Res. 2008;77(1):72-76.   DOI
23 Huang DT, Lu CY, Shao PL, Chang LY, Wang JY, Chang YH, Lai MJ, Chi YH, Huang LM. In vivo inhibition of influenza A virus replication by RNA interference targeting the PB2 subunit via intratracheal delivery. PLoS One. 2017;12(4):e0174523.   DOI
24 Gillman A, Muradrasoli S, Soderstrom H, Holmberg F, Latorre-Margalef N, Tolf C, Waldenstrom J, Gunnarsson G, Olsen B, Jarhult JD. Oseltamivir-resistant influenza A (H1N1) virus strain with an H274Y mutation in neuraminidase persists without drug pressure in infected mallards. Appl Environ Microbiol. 2015;81(7):2378-2383.   DOI
25 Govorkova EA, Baranovich T, Seiler P, Armstrong J, Burnham A, Guan Y, Peiris M, Webby RJ, Webster RG. Antiviral resistance among highly pathogenic influenza A (H5N1) viruses isolated worldwide in 2002- 2012 shows need for continued monitoring. Antiviral Res. 2013;98(2):297-304.   DOI
26 Fouchier RA, Bestebroer TM, Herfst S, Van Der Kemp L, Rimmelzwaan GF, Osterhaus AD. Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene. J Clin Microbiol. 2000;38(11):4096-4101.   DOI
27 Zhou H, Jin M, Yu Z, Xu X, Peng Y, Wu H, Liu J, Liu H, Cao S, Chen H. Effective small interfering RNAs targeting matrix and nucleocapsid protein gene inhibit influenza A virus replication in cells and mice. Antiviral Res. 2007;76(2):186-193.   DOI
28 Zhou K, He H, Wu Y, Duan M. RNA interference of avian influenza virus H5N1 by inhibiting viral mRNA with siRNA expression plasmids. J Biotechnol. 2008;135(2):140-144.   DOI
29 Dharmayanti NLPI, Ibrahim F, Soebandrio A. Amantadine resistant of Indonesian H5N1 subtype influenza viruses during 2003-2008. Microbiol Indones. 2010;4:1-6.   DOI
30 Lee MS, Chang PC, Shien JH, Cheng MC, Shieh HK. Identification and subtyping of avian influenza viruses by reverse transcription-PCR. J Virol Methods. 2001;97(1-2):13-22.   DOI
31 Stauber N, Brechtbuhl K, Bruckner L, Hofmann MA. Detection of Newcastle disease virus in poultry vaccines using the polymerase chain reaction and direct sequencing of amplified cDNA. Vaccine. 1995;13(4):360-364.   DOI
32 Wright KE, Wilson GA, Novosad D, Dimock C, Tan D, Weber JM. Typing and subtyping of influenza viruses in clinical samples by PCR. J Clin Microbiol. 1995;33(5):1180-1184.   DOI