Generation of an Immunostimulatory Protein-Encoding Recombinant Measles Virus

Begin with a liposome suspension encapsulating recombinant measles virus antigenome plasmid and helper plasmids encoding essential viral proteins.

The antigenome plasmid contains cDNA of the measles genome, modified to express immunostimulatory proteins and a fluorescent reporter in infected cells.

Take a virus producer cell culture containing antibiotics to prevent contamination.

Introduce the liposome suspension into the culture and incubate.

The liposomes fuse with the cell membrane to deliver the plasmids.

Host cell machinery transcribes the antigenome cDNA into RNA and the helper plasmids into mRNAs, enabling protein expression and formation of the viral replication complex. This complex produces genomic RNA, which enables viral protein expression.

Genomic RNA assembles with viral proteins to generate recombinant viral particles.

Infected cells also express the fluorescent reporter protein for microscopic visualization and produce the virus-encoded immunostimulatory protein.

Infected cells form multinucleated syncytia, a hallmark of measles' cytopathic effect.

Use fluorescence microscopy to identify large fluorescent syncytia, confirming successful recombinant virus rescue.

To rescue me