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Reverse genetics rescue of sylvatic dengue viruses
Paul Gendler1,2, Arturo Barbachano-Guerrero2, Samuel D Chappell2
1Department of Molecular, Cellular, and Developmental Biology, University of Colorado Boulder, Boulder, Colorado, USA.
Journal of Virology
|June 4, 2025
Summary
Researchers optimized a reverse genetics method to rescue sylvatic dengue viruses, enabling the study of forest-dwelling dengue virus strains and their potential for human adaptation. This advance provides crucial tools for understanding emerging infectious disease threats.
Area of Science:
- Virology
- Infectious Diseases
- Molecular Biology
Background:
- Sylvatic dengue viruses, circulating in nonhuman primates and mosquitoes in Africa and Asia, occasionally infect humans, posing a potential threat for new epidemics.
- A significant barrier to studying these viruses is the limited availability of infectious stocks, despite numerous genome sequences in public databases.
- Existing reverse genetics methods for dengue virus manipulation are challenging, particularly for sylvatic strains.
Purpose of the Study:
- To optimize a reverse genetics technique for the efficient rescue of infectious sylvatic dengue virus stocks from sequence data.
- To generate a collection of rescued sylvatic dengue virus strains for further biological characterization.
- To assess the replication fitness of different sylvatic dengue virus strains in various cell types, including human, monkey, and mosquito cells.
Main Methods:
- Development and optimization of a novel reverse genetics approach tailored for sylvatic dengue viruses.
- Utilizing mosquito cells, rather than mammalian cells, as the critical cell type for initiating sylvatic dengue virus replication from assembled genomes.
- Rescue of seven distinct sylvatic dengue virus strains using the optimized method.
Main Results:
- Successful rescue of seven sylvatic dengue virus strains, creating a valuable resource for research.
- Demonstration that mosquito cells are universally permissive for sylvatic dengue virus replication.
- Identification of specific sylvatic dengue virus strains exhibiting significantly enhanced replication in human and monkey cells compared to others.
Conclusions:
- The optimized reverse genetics method enables the transformation of any available sylvatic dengue virus sequence into infectious virus stocks.
- Certain sylvatic dengue virus strains show a higher replication potential in mammalian cells, suggesting a greater capacity for human adaptation.
- This work provides essential tools and insights for monitoring and understanding the potential emergence of novel dengue virus threats to human populations.
