ArticleProceedings of the National Academy of Sciences of the United States of America2014
Plasmid-based human norovirus reverse genetics system produces reporter-tagged progeny virus containing infectious genomic RNA.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
47 citing papers in PubMed, 75 citations in OpenAlex.
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- Conformational Plasticity of the Human Norovirus GII.3 Capsid Reveals Alternative P Domain Interaction Networks.International journal of molecular sciences · 2026Article
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- Overcoming host restrictions to enable continuous passaging of GII.3 human norovirus in human intestinal enteroids.Science advances · 2026Article
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- Recovery of infectious recombinant human norovirus using zebrafish embryos.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Current Perspectives and Future Directions in the Immunogenicity Landscape of Norovirus Vaccines.Journal of microbiology and biotechnology · 2025Review
- Plasmid-Based Reverse Genetics System Enabling One-Step Generation of Genotype 3 Hepatitis E Virus.Viruses · 2025Article
- Highs and Lows in Calicivirus Reverse Genetics.Viruses · 2024Review
- Human norovirus cultivation systems and their use in antiviral research.Journal of virology · 2024Review
- Production of infectious reporter murine norovirus by VP2Journal of virology · 2024Article
- Amino acid substitutions in norovirus VP1 dictate host dissemination via variations in cellular attachment.Journal of virology · 2023Article
- Trends on Human Norovirus Virus-like Particles (HuNoV-VLPs) and Strategies for the Construction of Infectious Viral Clones toward In Vitro Replication.Life (Basel, Switzerland) · 2023Review
- Molecular and Genetics-Based Systems for Tracing the Evolution and Exploring the Mechanisms of Human Norovirus Infections.International journal of molecular sciences · 2023Review
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- Recent insights into reverse genetics of norovirus.Virus research · 2023Review
- Experimental Adaptation of Murine Norovirus to Calcium Hydroxide.Frontiers in microbiology · 2022Article
- Advances in understanding of the innate immune response to human norovirus infection using organoid models.The Journal of general virology · 2022Review
- CD300lf Conditional Knockout Mouse Reveals Strain-Specific Cellular Tropism of Murine Norovirus.Journal of virology · 2021Article
Corrections and comments
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Authors and funding
10 authors at 4 institutions in 2 countries.
Funding
Abstract
Human norovirus (HuNoV) is the leading cause of gastroenteritis worldwide. HuNoV replication studies have been hampered by the inability to grow the virus in cultured cells. The HuNoV genome is a positive-sense single-stranded RNA (ssRNA) molecule with three open reading frames (ORFs). We established a reverse genetics system driven by a mammalian promoter that functions without helper virus. The complete genome of the HuNoV genogroup II.3 U201 strain was cloned downstream of an elongation factor-1α (EF-1α) mammalian promoter. Cells transfected with plasmid containing the full-length genome (pHuNoVU201F) expressed the ORF1 polyprotein, which was cleaved by the viral protease to produce the mature nonstructural viral proteins, and the capsid proteins. Progeny virus produced from the transfected cells contained the complete NoV genomic RNA (VP1, VP2, and VPg) and exhibited the same density in isopycnic cesium chloride gradients as native infectious NoV particles from a patient's stool. This system also was applied to drive murine NoV RNA replication and produced infectious progeny virions. A GFP reporter construct containing the GFP gene in ORF1 produced complete virions that contain VPg-linked RNA. RNA from virions containing the encapsidated GFP-genomic RNA was successfully transfected back into cells producing fluorescent puncta, indicating that the encapsidated RNA is replication-competent. The EF-1α mammalian promoter expression system provides the first reverse genetics system, to our knowledge, generalizable for human and animal NoVs that does not require a helper virus. Establishing a complete reverse genetics system expressed from cDNA for HuNoVs now allows the manipulation of the viral genome and production of reporter virions.
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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.