Recombinant Rhabdovirus containing a heterologous fusion protein
AssigneeThe University of Tennessee Research Corporation
InventorsMichael A. Whitt, Clinton S. Robison
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Abstract
This invention relates to a composition comprising a recombinant or genetically engineered Rhabdovirus that expresses a Fusion Protein, such as the F protein of the Paramyxovirus SV5 strain. This recombinant Rhabdovirus may express other non-Rhabdovirus attachment proteins and/or an enhancer protein. The invention also relates to methods of making recombinant Rhabdoviruses which express an F Protein. These recombinant compositions can be used for purposes of research, as well as for diagnostic and therapeutic compositions for treatment of diseases.
Claims28
A genetically engineered Rhabdovirus comprising:
- a heterologous fusion protein, that is effective to facilitate fusion of the Rhabdovirus to a target cell membrane;
- a heterologous G stem polypeptide;
- the Rhabdovirus N, P and L proteins;
- and a deletion of the nucleic acid sequence which encodes G protein of the Rhabdovirus.
A method of producing a recombinant Rhabdovirus comprising an F Protein effective to facilitate fusion of the Rhabdovirus to a cell membrane thereof and a heterologous G stem protein comprising the steps of:
- (A) inserting a cDNA encoding Rhabdovirus N, P, L and G proteins into a suitable cell;
- (B) inserting a polycistronic cDNA copy of the Rhabdovirus genome containing at least the 3′ and 5′ Rhabdovirus leader and trailer regions containing the cis acting signals for Rhabdovirus replication, the genes encoding the N, P, M, and L Rhabdovirus proteins and a gene encoding an F Protein, wherein the nucleic acid sequence which encodes G protein of the Rhabdovirus is deleted and a heterologous G stem protein into the suitable cell;
- (C) culturing the cell under conditions that permit production of the recombinant Rhabdovirus;
- and (D) isolating said recombinant Rhabdovirus.
A method of producing a recombinant Rhabdovirus which expresses an F Protein thereof effective to facilitate fusion of the Rhabdovirus to a cell membrane comprising the steps of:
- (A) inserting into suitable cells a polycistronic cDNA comprising at least the 3′ and 5′ Rhabdovirus leader and trailer regions containing the cis acting signals for Rhabdovirus replication, the genes encoding the Rhabdovirus N, P, and L proteins and a gene encoding an F Protein;
- (B) infecting the cells with a minivirus comprising the cis acting signals for Rhabdovirus replication and genes encoding the Rhabdovirus M protein and a heterologous G stem polypeptide, wherein the nucleic acid sequence which encodes G protein of the Rhabdovirus is deleted;
- (C) culturing the cells under conditions to permit expression of the cDNA to produce the recombinant Rhabdovirus;
- and
- (D) isolating said recombinant Rhabdovirus.
A recombinant Rhabdovirus comprising a Rhabdovirus genome wherein the Rhabdovirus comprises a deletion of the nucleic acid sequence which encodes a G protein gene of the Rhabdovirus, and a nucleic acid which encodes a heterologous fusion polypeptide, and a nucleic acid which encodes a heterologous G stem polypeptide.
Dependencies are inferred from the claim text.