ArticlePloS one2016
Enterovirus A71 DNA-Launched Infectious Clone as a Robust Reverse Genetic Tool.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 43 citations in OpenAlex.
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- Codon-deoptimized single-round infectious virus for therapeutic and vaccine applications.Scientific reports · 2025Article
- Novel reporter constructs to accelerate antiviral and therapeutic discovery for Enterovirus-A71.Antiviral research · 2025Article
- Exploring enterovirus pathogenesis and cancer therapy potential through reverse genetics.Biosafety and health · 2025Review
- Functional anatomy of zinc finger antiviral protein complexes.Nature communications · 2024Article
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- Enterovirus 3A protein disrupts endoplasmic reticulum homeostasis through interaction with GBF1.Journal of virology · 2024Article
- Development and validation of Mayaro virus with luciferase reporter genes as a tool for antiviral assays.Heliyon · 2024Article
- Construction of an infectious clone for enterovirus A89 and mutagenesis analysis of viral infection and cell binding.Microbiology spectrum · 2024Article
- Substitution of Coxsackievirus A16 VP1 BC and EF Loop Altered the Protective Immune Responses in Chimera Enterovirus A71.Vaccines · 2023Article
- Engineering Oncolytic Coxsackievirus A21 with Small Transgenes and Enabling Cell-Mediated Virus Delivery by Integrating Viral cDNA into the Genome.Journal of virology · 2023Article
- Development of a Universal Cloning System for Reverse Genetics of Human Enteroviruses.Microbiology spectrum · 2023Article
- Pharmacological perturbation of CXCL1 signaling alleviates neuropathogenesis in a model of HEVA71 infection.Nature communications · 2022Article
- Procedures and potential pitfalls for constructing a bee-infecting RNA virus clone.Frontiers in insect science · 2022Review
- Isolation and Identification of Inter-SpeciesViruses · 2021Article
- A versatile reverse genetics platform for SARS-CoV-2 and other positive-strand RNA viruses.Nature communications · 2021Article
- Recovery and Genetic Characterization of a West Nile Virus Isolate from China.Virologica Sinica · 2021Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Enterovirus A71 (EV-A71) causes major outbreaks of hand, foot and mouth disease, and is occasionally associated with neurological complications and death in children. Reverse genetics is widely used in the field of virology for functional study of viral genes. For EV-A71, such tools are limited to clones that are transcriptionally controlled by T7/SP6 bacteriophage promoter. This is often time-consuming and expensive. Here, we describe the development of infectious plasmid DNA-based EV-A71 clones, for which EV-A71 genome expression is under transcriptional control by the CMV-intermediate early promoter and SV40 transcriptional-termination signal. Transfection of this EV-A71 infectious DNA produces good virus yield similar to in vitro-transcribed EV-A71 infectious RNA, 6.4 and 5.8 log10PFU/ml, respectively. Infectious plasmid with enhanced green fluorescence protein and Nano luciferase reporter genes also produced good virus titers, with 4.3 and 5.0 log10 PFU/ml, respectively. Another infectious plasmid with both CMV and T7 promoters was also developed for easy manipulation of in vitro transcription or direct plasmid transfection. Transfection with either dual-promoter infectious plasmid DNA or infectious RNA derived from this dual-promoter clone produced infectious viral particles. Incorporation of hepatitis delta virus ribozyme, which yields precise 3' ends of the DNA-launched EV-A71 genomic transcripts, increased infectious viral production. In contrast, the incorporation of hammerhead ribozyme in the DNA-launched EV-A71 resulted in lower virus yield, but improved the virus titers for T7 promoter-derived infectious RNA. This study describes rapid and robust reverse genetic tools for EV-A71.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.