Key result
A real-time fluorescence-based protocol using SYBR Green II and RNase was developed to detect the viral genome during various stages of enterovirus uncoating in vitro.
Why the study?
Cues leading to enterovirus uncoating are less well known, and monitoring has been limited by a lack of methods detecting dynamic virion changes.
A novel real-time fluorescence protocol enables the dynamic monitoring of enterovirus uncoating in vitro.
Provides novel assay for enterovirus uncoating dynamics; leaves open physiological relevance and requires cellular/in vivo validation.
Viruses need to open, i.e., uncoat, in order to release their genomes for efficient replication and translation. Especially for non-enveloped viruses, such as enteroviruses, the cues leading to uncoating are less well known. The status of the virus has previously been observed mainly by transmission electron microscopy using negative staining, cryo electron microscopy, X-ray crystallography or gradient separation (reviewed in However, monitoring of uncoating has been limited by the lack of methods detecting dynamic changes of the virions. Here, we present a real-time fluorescence based protocol, which detects the viral genome (RNA) during various stages of uncoating in vitro, while RNA is still inside the particle that has been expanded before the actual RNA release, and when the RNA has been totally released from the viral particle. Our method allows to explore how various molecular factors may promote or inhibit virus uncoating.
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Ruokolainen et al. (2020) studied Enterovirus uncoating (in vitro). Real-time fluorescence measurement protocol was evaluated. A real-time fluorescence-based protocol using SYBR Green II and RNase was developed to detect the viral genome during various stages of enterovirus uncoating in vitro.
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