Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Mar;23(2):e24.
doi: 10.4142/jvs.21174. Epub 2022 Jan 10.

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

Risza Hartawan  1   2 , Dwi Ari Pujianto  3 , Ni Luh Putu Indi Dharmayanti  4 , Amin Soebandrio  5
Affiliations

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

Risza Hartawan et al. J Vet Sci. 2022 Mar.

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: H5N1 subtype; Indonesia; RNA interference; antiviral agents; influenza in birds.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1. siRNA design for the nucleoprotein gene of the Indonesian H5N1 viruses using the siDirect Version 2.0 software. Two selected siRNA sequences were marked by blue color.
siRNA, small interfering RNA.
Fig. 2
Fig. 2. Relative expression of the H5N1 genes in the Marbin-Darby canine kidney cell as effects of the siRNA-NPs inhibition, including NP (A), PA (B), H5 (C), N1 (D), M (E), and NS (F). The differences between groups were analyzed with one-way ANOVA post hoc Bonferroni.
siRNA-NP, small interfering RNA design targeting the nucleoprotein gene; NP, nucleoprotein; PA, polymerase acidic; H5, hemagglutinin; N1, neuraminidase; M, matrix; NS, non-structural. *p < 0.05, †p < 0.01.
Fig. 3
Fig. 3. Mutation analysis of the nucleoprotein gene as the effect of three serial exposure of siRNA-NPs. The difference between the siRNA-NP and its target sequence was shown by bar marking. The mutations that occurred after siRNA-NP exposures are indicated by the arrows.
siRNA-NP, small interfering RNA design targeting the nucleoprotein gene.
See this image and copyright information in PMC

References

    1. Sims LD, Brown IH. In: Animal Influenza. 2nd ed. Swayne DE, editor. Iowa: John Wiley & Sons, Inc.; 2016. Multi-continental panzootic of H5 highly pathogenic avian influenza (1996–2015) pp. 204–247.
    1. 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. - PMC - PubMed
    1. Shaw ML, Palese P. In: Fields Virology. 6th ed. Knipe DM, Howley PM, editors. Baltimore: Wolter Luwer; 2013. Orthomyxoviridae; pp. 1151–1185.
    1. Suarez DL. In: Animal Influenza. 2nd ed. Swayne DE, editor. Iowa: John Wiley & Sons, Inc.; 2016. Influenza A virus; pp. 3–30.
    1. 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. - PMC - PubMed