Transgenic expression of PB2 in 293T cells increases avian influenza virus packaging in reverse genetic systems

Transgenic expression of PB2 in 293T cells increases avian influenza virus packaging in reverse genetic systems | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 31 January 2024 V1 Latest version Share on Transgenic expression of PB2 in 293T cells increases avian influenza virus packaging in reverse genetic systems Authors : ying Xie 0000-0001-5147-874X , Ying Cui , Xinghai Zhang , Yuanguo Li , Weiyang Sun , Na Feng , Tiecheng Wang , Yongkun Zhao , Songtao Yang , Yuwei Gao , Chuan Qin , and Xianzhu Xia [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.170671023.38851889/v1 438 views 138 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Plasmid-based reverse genetics has transformed influenza virus research by enabling the production of recombinant influenza viruses from cloned cDNA copies of viral genome segments. Reverse genetic production of influenza viruses requires cellular expression of influenza proteins polymerase basic 1, polymerase basic 2, polymerase acidic and nucleoprotein which collectively allow for transcription of viral mRNA and synthesis of new negative-sense genomic RNA, thus enabling synthesis of all components needed to assemble infectious virus from transfected cell lines. Given the importance of these proteins in the generation of influenza viruses via reverse genetics, we sought to explore how transgenic expression of mammalian-adapted PB1, PB2, PA, or NP in the 293T packaging cell line may impact the recovery of recombinant influenza viruses. We constructed four transgenic 293T cell lines expressing PB1, PB2, PA or NP derived from the mouse-adapted 2009 pandemic influenza A virus, UI182. Transgenic expression of UI182 PB2 in 293T cells enhanced recovery of replication-competent avian influenza viruses generated by reverse genetics relative to levels achieved in unmodified 293T cells. Virus recovered from PB2-expressing 293T cells replicated with kinetics that were indistinguishable from viruses recovered from unmodified 293T cells. Provision of UI182 PB2 protein via transgenic expression in 293T cells resulted in enhanced viral polymerase activity as measured by a minigenome assay, which may account for the improved efficiency of viral packaging relative to unmodified 293T cells. Transgenic expression of mammalian-adapted PB2 in 293T cells may serve as an important tool for enhancing influenza virus recovery in reverse genetic systems. Supplementary Material File (manuscript.doc) Download 260.50 KB Information & Authors Information Version history V1 Version 1 31 January 2024 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords influenza a polymerase basic 2 reverse genetics transgenic 293t cells Authors Affiliations ying Xie 0000-0001-5147-874X Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center View all articles by this author Ying Cui Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Xinghai Zhang Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Yuanguo Li Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Weiyang Sun Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Na Feng Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Tiecheng Wang Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Yongkun Zhao Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Songtao Yang Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Yuwei Gao Military Veterinary Research Institute of Academy of Military Medical Sciences View all articles by this author Chuan Qin Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center View all articles by this author Xianzhu Xia [email protected] Chinese Academy of Medical Sciences (CAMS) and Comparative Medicine Center View all articles by this author Metrics & Citations Metrics Article Usage 438 views 138 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation ying Xie, Ying Cui, Xinghai Zhang, et al. Transgenic expression of PB2 in 293T cells increases avian influenza virus packaging in reverse genetic systems. Authorea . 31 January 2024. DOI: https://doi.org/10.22541/au.170671023.38851889/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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