Co-Packaging of TBEV Protease and Minigenome in FLAG Tagged Virus Like Particles and Extracellular Vesicles

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22330%2F25%3A43932652" target="_blank" >RIV/60461373:22330/25:43932652 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Co-Packaging of TBEV Protease and Minigenome in FLAG Tagged Virus Like Particles and Extracellular Vesicles

Popis výsledku v původním jazyce

Tick‑borne encephalitis virus (TBEV) is a positive‑sense RNA virus belonging to the flavivirus genus: It is the causative agent of tick‑borne encephalitis, which is a serious disease affecting the central nervous system.Virus‑like particles (VLPs) derived from TBEV represent a promising tool for studying viral assembly and RNA packaging due to their structural similarity to native virions and ability to encapsulate genetic material withoutbeing infectious. We engineered TBEV VLPs by expressing TBEV structural proteins: capsid (C), premembrane (prM), and envelope (E) proteins, in mammalian cells. To facilitate detection and tracking, we inserted a FLAG‑tag into the C protein of these VLPs. We generated a series of TBEV‑derived minigenomes of varying lengths and sequence compositions, focusing on the presence or absence of untranslated regions (UTRs). Our results demonstrate that minigenomes containing an intact 3’ UTR or 5’ UTR are incorporated into VLPs more efficiently than truncated variants lacking these UTRs or those extended with both UTRs. This system enables us to study the critical role of UTRs in selective RNA packaging, providing a versatile system to dissect the contributions of specific genomic elements to RNA encapsidation.Additionally, we conducted a detailed analysis of the packaging and intracellular co‑localization of TBEV‑derived minigenomes, together with the protease complex (NS2B–NS3) or TBEV structural proteins (C, prM, E) within extracellular vesicles (EVs) secreted by producer cells. The observed association of viral components with EVs suggests a potential pathway for RNA transport and intercellular communication that may contribute to viral dissemination or immune modulation.This work was supported by the project National Institute of Virology and Bacteriology (Programme EXCELES, ID Project No. LX22NPO5103) – Funded by the European Union – Next Generation EU.

Název v anglickém jazyce

Co-Packaging of TBEV Protease and Minigenome in FLAG Tagged Virus Like Particles and Extracellular Vesicles

Popis výsledku anglicky

Tick‑borne encephalitis virus (TBEV) is a positive‑sense RNA virus belonging to the flavivirus genus: It is the causative agent of tick‑borne encephalitis, which is a serious disease affecting the central nervous system.Virus‑like particles (VLPs) derived from TBEV represent a promising tool for studying viral assembly and RNA packaging due to their structural similarity to native virions and ability to encapsulate genetic material withoutbeing infectious. We engineered TBEV VLPs by expressing TBEV structural proteins: capsid (C), premembrane (prM), and envelope (E) proteins, in mammalian cells. To facilitate detection and tracking, we inserted a FLAG‑tag into the C protein of these VLPs. We generated a series of TBEV‑derived minigenomes of varying lengths and sequence compositions, focusing on the presence or absence of untranslated regions (UTRs). Our results demonstrate that minigenomes containing an intact 3’ UTR or 5’ UTR are incorporated into VLPs more efficiently than truncated variants lacking these UTRs or those extended with both UTRs. This system enables us to study the critical role of UTRs in selective RNA packaging, providing a versatile system to dissect the contributions of specific genomic elements to RNA encapsidation.Additionally, we conducted a detailed analysis of the packaging and intracellular co‑localization of TBEV‑derived minigenomes, together with the protease complex (NS2B–NS3) or TBEV structural proteins (C, prM, E) within extracellular vesicles (EVs) secreted by producer cells. The observed association of viral components with EVs suggests a potential pathway for RNA transport and intercellular communication that may contribute to viral dissemination or immune modulation.This work was supported by the project National Institute of Virology and Bacteriology (Programme EXCELES, ID Project No. LX22NPO5103) – Funded by the European Union – Next Generation EU.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10607 - Virology

Projekt

<a href="/cs/project/LX22NPO5103" target="_blank" >LX22NPO5103: Národní institut virologie a bakteriologie</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů