Direct protein delivery with engineered microvesicles
View Patent ↗The present invention relates to direct protein delivery with engineered micro vesicles.
1. An in vitro method of delivering a protein of interest into a target cell comprising culturing a target cell with an isolated eukaryotic cell overexpressing a viral membrane fusion protein and a protein of interest, wherein said eukaryotic cell secretes microvesicles comprising said viral membrane fusion protein and a protein of interest, wherein said eukaryotic cell does not express any viral structural proteins and said protein of interest is not covalently bonded to said viral membrane fusion protein, and wherein said microvesicles secreted by said eukaryotic cell contact said target cell and deliver said protein of interest into said target cell.
2. The method according to claim 1 , wherein several proteins of interest are delivered into said target cell by culturing said target cell with several of said eukaryotic cell each overexpressing a different protein of interest.
3. The method according to claim 1 , wherein said protein of interest is a membrane receptor.
4. The method according to claim 1 , wherein said protein of interest is a transcription factor.
5. A method of rendering a target cell transiently permissive to an ecotropic viral particle comprising contacting a target cell with a microvesicle comprising a viral fusion protein and an ecotrophic viral receptor (EcoR), wherein said contacting delivers said EcoR into said target cell and renders said target cell transiently permissive to an ecotropic viral particle, wherein said microvesicle is produced by an isolated eukaryotic cell overexpressing a viral membrane fusion protein and said EcoR, wherein said eukaryotic cell secretes said microvesicles, and wherein said eukaryotic cell does not express any viral structural proteins and said EcoR is not covalently bonded to said viral membrane fusion protein.