Nucleocapsid-independent specific viral RNA packaging and uses thereof
OwnerShinji Makino, Krishna Narayananinventors
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Abstract
The present invention shows that expressed coronavirus envelope protein M specifically interacted with co-expressed non-coronavirus RNA transcripts containing the short viral packaging signal in the absence of coronavirus N protein. Furthermore, this M protein-packaging signal interaction led to specific packaging of the packaging-signal-containing RNA transcripts into coronavirus-like particles in the absence of N protein. These findings highlight a novel RNA packaging mechanism for an enveloped virus, and a novel coronavirus-based expression system can be developed based on the data presented herein.
Claims10
A coronavirus-based gene delivery and expression vector system comprising:
- a vector comprising a promoter, a sequence encoding a protein of interest and a packaging signal of a coronavirus;
- and
- a vector or vectors encoding the envelope protein(s) of a coronavirus.
A method of expressing a protein of interest in target cells, said method comprises the steps of:
- transfecting a first set of cells with a vector, said vector comprises a promoter, a sequence encoding a protein of interest and a packaging signal of a coronavirus;
- transcribing said sequence encoding said protein of interest into RNA molecules;
- transfecting or infecting said first cells with a vector or vectors encoding one or more of the envelope proteins of a coronavirus;
- packaging said RNA molecules and said envelope protein(s) into viral particles;
- collecting said viral particles comprising said RNA molecules encoding said protein of interest;
- and
- infecting target cells with said viral particles, wherein translation of said RNA molecules transferred to said target cells by said viral particles results in expression of said protein of interest in said target cells.
Dependencies are inferred from the claim text.