JoVE Encyclopedia of Experiments
Microbiology
0 views • 3:08 min • September 30th, 2026
Start with a minigenome plasmid that mimics the Ebola virus genome and encodes viral structural proteins, glycoproteins, and luciferase reporter.
Add helper plasmids that encode the T7 polymerase, viral nucleoprotein, and viral polymerase components essential for replication and transcription.
Add a lipid-based transfection reagent to the plasmids, vortex, and incubate the mixture to form plasmid-lipid complexes.
Add this mixture to human kidney cells, mix gently, and incubate.
The lipid complex fuses with the cell. Plasmids enter the cell.
The helper plasmids encode their respective proteins.
Later, T7 polymerase transcribes the minigenome into viral genomic RNA, which is immediately coated with nucleoproteins.
The viral polymerase complex binds to the viral genomic RNA and transcribes it into viral mRNAs. These mRNAs produce viral structural proteins and glycoproteins.
The viral proteins assemble to form non-pathogenic transcription-and replication-competent virus-like particles or trVLPs containing the luciferase reporter gene.
Replenish with fresh medium to maintain cell viability and support trVLP production.
Transfect the cells with the plasmids required for production of TRVLPs. In a
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