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Poult Sci . Research Note: Establishment of vector system harboring duck RNA polymerase I promoter for avian influenza virus

tetano

Editor, Senior Moderator
Poult Sci


. 2024 Nov 22;104(1):104570.
doi: 10.1016/j.psj.2024.104570. Online ahead of print. Research Note: Establishment of vector system harboring duck RNA polymerase I promoter for avian influenza virus

Yunyueng Jang[SUP] 1 [/SUP], Yoon-Gi Baek[SUP] 1 [/SUP], Yu-Na Lee[SUP] 1 [/SUP], Ra Mi Cha[SUP] 1 [/SUP], Yun-Chan Choi[SUP] 1 [/SUP], Min-Ji Park[SUP] 1 [/SUP], Youn-Jeong Lee[SUP] 1 [/SUP], Eun-Kyoung Lee[SUP] 2 [/SUP]



Affiliations
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.

Keywords: Avian influenza; Duck RNA polymerase I; Reverse genetics.

 
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