Research Note: Establishment of Vector System Harboring Duck RNA Polymerase I Promoter for Avian Influenza Virus

Overview

Journal Poult Sci

Publisher Elsevier

Specialty Veterinary Medicine

Date 2024 Dec 4

PMID 39631283

Authors

Yunyueng Jang

Yoon-Gi Baek

Yu-Na Lee

Ra Mi Cha

Yun-Chan Choi

Min-Ji Park

Youn-Jeong Lee

Eun-Kyoung Lee

Affiliations

Soon will be listed here.

Abstract

Reverse genetics (RG) systems are extensively utilized to investigate the characteristics of influenza viruses and develop vaccines, predominantly relying on human RNA polymerase I (pol I). However, the efficiency of RG systems for avian-origin influenza viruses may be compromised due to potential species-specific interactions of RNA pol I. In this study, we reported the polymerase activities of the duck RNA pol I promoter in avian cells and the generation of recombinant avian-derived influenza viruses using a novel vector system containing the duck RNA pol I promoter region to enhance the rescue efficiency of the RG system in avian cells. Initially, we explored a putative duck promoter region and identified the optimal size to improve the existing system. Subsequently, we established an RG system incorporating the duck RNA pol I promoter and compared its rescue efficiency with the human pol I system by generating recombinant influenza viruses in several cell lines. Notably, the 250-bp duck RNA pol I promoter demonstrated effective functionality in avian cells, exhibiting higher polymerase activity in a minigenome assay. The newly constructed RG system was significantly improved, enabling the rescue of influenza viruses in 293T, DF-1, and CCL141 cells. Furthermore, HPAI viruses were successfully rescued in DF-1 cells, a result that had not been achieved in previous experiments. In conclusion, our novel RG system harboring duck RNA pol I offers an additional tool for researching influenza viruses and may facilitate the development of vaccines for poultry.

References

Suphaphiphat P, Keiner B, Trusheim H, Crotta S, Tuccino A, Zhang P . Human RNA polymerase I-driven reverse genetics for influenza a virus in canine cells. J Virol. 2010; 84(7):3721-5. PMC: 2838141. DOI: 10.1128/JVI.01925-09. View

Hoffmann E, Neumann G, Kawaoka Y, HOBOM G, Webster R . A DNA transfection system for generation of influenza A virus from eight plasmids. Proc Natl Acad Sci U S A. 2000; 97(11):6108-13. PMC: 18566. DOI: 10.1073/pnas.100133697. View

Liu L, Madhugiri R, Saul V, Bacher S, Kracht M, Pleschka S . Phosphorylation of the PA subunit of influenza polymerase at Y393 prevents binding of the 5'-termini of RNA and polymerase function. Sci Rep. 2023; 13(1):7042. PMC: 10148841. DOI: 10.1038/s41598-023-34285-7. View

Song M, Baek Y, Pascua P, Kwon H, Park S, Kim E . Establishment of Vero cell RNA polymerase I-driven reverse genetics for Influenza A virus and its application for pandemic (H1N1) 2009 influenza virus vaccine production. J Gen Virol. 2013; 94(Pt 6):1230-1235. DOI: 10.1099/vir.0.051284-0. View

Nogales A, Martinez-Sobrido L . Reverse Genetics Approaches for the Development of Influenza Vaccines. Int J Mol Sci. 2016; 18(1). PMC: 5297655. DOI: 10.3390/ijms18010020. View

Shinagawa T, Ishii S . Generation of Ski-knockdown mice by expressing a long double-strand RNA from an RNA polymerase II promoter. Genes Dev. 2003; 17(11):1340-5. PMC: 196066. DOI: 10.1101/gad.1073003. View

Diebel K, Smith A, van Dyk L . Mature and functional viral miRNAs transcribed from novel RNA polymerase III promoters. RNA. 2009; 16(1):170-85. PMC: 2802027. DOI: 10.1261/rna.1873910. View

Paule M, White R . Survey and summary: transcription by RNA polymerases I and III. Nucleic Acids Res. 2000; 28(6):1283-98. PMC: 111039. DOI: 10.1093/nar/28.6.1283. View

Heix J, Grummt I . Species specificity of transcription by RNA polymerase I. Curr Opin Genet Dev. 1995; 5(5):652-6. DOI: 10.1016/0959-437x(95)80035-2. View

10.

Cha R, Lee Y, Park M, Baek Y, Shin J, Jung C . Genetic Characterization and Pathogenesis of H5N1 High Pathogenicity Avian Influenza Virus Isolated in South Korea during 2021-2022. Viruses. 2023; 15(6). PMC: 10304347. DOI: 10.3390/v15061403. View

11.

Dou D, Revol R, Ostbye H, Wang H, Daniels R . Influenza A Virus Cell Entry, Replication, Virion Assembly and Movement. Front Immunol. 2018; 9:1581. PMC: 6062596. DOI: 10.3389/fimmu.2018.01581. View

12.

Murakami S, Horimoto T, Yamada S, Kakugawa S, Goto H, Kawaoka Y . Establishment of canine RNA polymerase I-driven reverse genetics for influenza A virus: its application for H5N1 vaccine production. J Virol. 2007; 82(3):1605-9. PMC: 2224466. DOI: 10.1128/JVI.01876-07. View

13.

Li O, Chan M, Leung C, Chan R, Guan Y, Nicholls J . Full factorial analysis of mammalian and avian influenza polymerase subunits suggests a role of an efficient polymerase for virus adaptation. PLoS One. 2009; 4(5):e5658. PMC: 2680953. DOI: 10.1371/journal.pone.0005658. View

14.

Suzuki Y, Uchida Y, Tanikawa T, Maeda N, Takemae N, Saito T . Amino acid substitutions in PB1 of avian influenza viruses influence pathogenicity and transmissibility in chickens. J Virol. 2014; 88(19):11130-9. PMC: 4178826. DOI: 10.1128/JVI.01564-14. View

15.

Neumann G, Fujii K, Kino Y, Kawaoka Y . An improved reverse genetics system for influenza A virus generation and its implications for vaccine production. Proc Natl Acad Sci U S A. 2005; 102(46):16825-9. PMC: 1283806. DOI: 10.1073/pnas.0505587102. View

16.

Wang K, Huang Q, Yang Z, Qi K, Liu H, Chen H . Alternative reverse genetics system for influenza viruses based on a synthesized swine 45S rRNA promoter. Virus Genes. 2017; 53(4):661-666. DOI: 10.1007/s11262-017-1457-8. View

17.

Wang Z, Duke G . Cloning of the canine RNA polymerase I promoter and establishment of reverse genetics for influenza A and B in MDCK cells. Virol J. 2007; 4:102. PMC: 2241602. DOI: 10.1186/1743-422X-4-102. View

18.

Moncorge O, Long J, Cauldwell A, Zhou H, Lycett S, Barclay W . Investigation of influenza virus polymerase activity in pig cells. J Virol. 2012; 87(1):384-94. PMC: 3536367. DOI: 10.1128/JVI.01633-12. View

19.

Lee C, Jung K, Jadhao S, Suarez D . Evaluation of chicken-origin (DF-1) and quail-origin (QT-6) fibroblast cell lines for replication of avian influenza viruses. J Virol Methods. 2008; 153(1):22-8. DOI: 10.1016/j.jviromet.2008.06.019. View

20.

Grummt I . Life on a planet of its own: regulation of RNA polymerase I transcription in the nucleolus. Genes Dev. 2003; 17(14):1691-702. DOI: 10.1101/gad.1098503R. View