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Updated: Jan 4, 2026
Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone
Published on: June 7, 2017
The Rescue and Characterization of Recombinant, Microcephaly-Associated Zika Viruses as Single-Round Infectious
Chien-Yi Lu1, Chen-Sheng Lin2, Hsueh-Chou Lai3,4
1Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung 404, Taiwan. cylu0424@gmail.com.
Abstract:
Zika virus (ZIKV) is transmitted by Aedes mosquitoes and exhibits genetic variation with African and Asian lineages. ZIKV Natal RGN strain, an Asian-lineage virus, has been identified in brain tissues from fetal autopsy cases with microcephaly and is suggested to be a neurotropic variant. However, ZIKV Natal RGN strain has not been isolated; its biological features are not yet illustrated. This study rescued and characterized recombinant, single-round infectious particles (SRIPs) of the ZIKV Natal RGN strain using reverse genetic and synthetic biology techniques. The DNA-launched replicon of ZIKV Natal RGN was constructed and contains the EGFP reporter, lacks prM-E genes, and replicates under CMV promoter control. The peak in the ZIKV Natal RGN SRIP titer reached 6.25 × 106 TCID50/mL in the supernatant of prM-E-expressing packaging cells 72 h post-transfection with a ZIKV Natal RGN replicon. The infectivity of ZIKV Natal RGN SRIPs has been demonstrated to correlate with the green florescence intensity of the EGFP reporter, the SRIP-induced cytopathic effect, and ZIKV's non-structural protein expression. Moreover, ZIKV Natal RGN SRIPs effectively self-replicated in rhabdomyosarcoma/muscle, glioblastoma/astrocytoma, and retinal pigmented epithelial cells, displaying unique cell susceptibility with differential attachment activity. Therefore, the recombinant ZIKV Natal RGN strain was rescued as SRIPs that could be used to elucidate the biological features of a neurotropic strain regarding cell tropism and pathogenic components, apply for antiviral agent screening, and develop vaccine candidates.
Insights
Researchers rescued and characterized Zika virus (ZIKV) Natal RGN strain as single-round infectious particles (SRIPs). These SRIPs enable study of neurotropic ZIKV variants, aiding antiviral and vaccine development.
Area of Science:
- Virology
- Neuroscience
- Synthetic Biology
Background:
- Zika virus (ZIKV) is transmitted by Aedes mosquitoes and has African and Asian lineages.
- The ZIKV Natal RGN strain, an Asian lineage, is linked to microcephaly and suspected neurotropism.
- The biological characteristics of the ZIKV Natal RGN strain remain largely unillustrated due to lack of isolation.
Purpose of the Study:
- To rescue and characterize recombinant single-round infectious particles (SRIPs) of the ZIKV Natal RGN strain.
- To elucidate the biological features of this putatively neurotropic ZIKV strain.
- To establish a tool for antiviral screening and vaccine development.
Main Methods:
- Utilized reverse genetics and synthetic biology to construct a DNA-launched ZIKV Natal RGN replicon with an EGFP reporter.
- Produced ZIKV Natal RGN SRIPs in packaging cells expressing prM-E genes.
- Assessed SRIP infectivity via EGFP expression, cytopathic effects, and non-structural protein expression.
Main Results:
- Successfully rescued ZIKV Natal RGN SRIPs with a peak titer of 6.25 × 10^6 TCID50/mL.
- Demonstrated SRIP infectivity correlated with EGFP intensity, cytopathic effects, and viral protein expression.
- Showed ZIKV Natal RGN SRIPs self-replicated in various cell types (muscle, glioblastoma, retinal), indicating unique cell tropism.
Conclusions:
- Recombinant ZIKV Natal RGN SRIPs were successfully generated, providing a viable model for studying neurotropic ZIKV.
- The characterized SRIPs facilitate investigation into ZIKV cell tropism and pathogenic mechanisms.
- These SRIPs serve as a valuable platform for screening antiviral agents and developing ZIKV vaccines.
