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Replication-Independent Generation and Morphological Analysis of Flavivirus Replication Organelles.


ABSTRACT: Positive-strand RNA viruses replicate in distinct membranous structures called replication organelles (ROs). Mechanistic studies of RO formation have been difficult because perturbations affecting viral replication have an impact on viral protein amounts, thus affecting RO biogenesis. Here, we present a detailed guide on how to use a replication-independent expression system, designated pIRO (plasmid-induced replication organelle formation), inducing bona fide flavivirus ROs in transfected cells. This will be useful for mechanistic studies of viral and cellular factors driving flavivirus RO biogenesis. For complete details on the use and execution of this protocol, please refer to Cerikan et al. (2020).

SUBMITTER: Goellner S 

PROVIDER: S-EPMC7757399 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Replication-Independent Generation and Morphological Analysis of Flavivirus Replication Organelles.

Goellner Sarah S   Cerikan Berati B   Cortese Mirko M   Neufeldt Christopher J CJ   Haselmann Uta U   Bartenschlager Ralf R  

STAR protocols 20201125 3


Positive-strand RNA viruses replicate in distinct membranous structures called replication organelles (ROs). Mechanistic studies of RO formation have been difficult because perturbations affecting viral replication have an impact on viral protein amounts, thus affecting RO biogenesis. Here, we present a detailed guide on how to use a replication-independent expression system, designated pIRO (plasmid-induced replication organelle formation), inducing bona fide flavivirus ROs in transfected cells  ...[more]

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