Generation of Reporter Gene-Carrying Hepatitis B Pseudoviruses

Take a mixture of plasmids encoding hepatitis B surface antigens and lentiviral proteins, along with a plasmid containing a fluorescent reporter gene and a packaging signal.

Add a transfection reagent, mix thoroughly, and incubate.

The positively charged transfection reagent forms complexes with the negatively charged plasmid DNA.

Introduce these complexes to a monolayer of human embryonic kidney cells and incubate.

The complexes enter the cells via endocytosis.

Replace the medium to remove residual complexes and incubate again.

Inside the cells, the plasmids produce hepatitis B surface antigens, lentiviral proteins, and RNA carrying the reporter sequence and packaging signal.

The packaging signal allows the reporter RNA to be packaged into the viral particles.

The assembled viral structures bud off as pseudoviruses displaying hepatitis B surface antigens.

Harvest the medium and centrifuge to remove cellular debris.

Collect the supernatant containing Hepatitis B pseudoviruses carrying the fluorescent reporter gene.