Replication-defective arenavirus vectors
The invention relates to an infectious arenavirus particle that is engineered to contain a genome with the ability to amplify and express its genetic information in infected cells but unable to produce further infectious progeny particles in normal, not genetically engineered cells. One or more of the four arenavirus open reading frames glycoprotein (GP), nucleoprotein (NP), matrix protein Z and RNA-dependent RNA polymerase L are removed or mutated to prevent replication in normal cells but still allowing gene expression in arenavirus vector-infected cells, and foreign genes coding for an antigen or other protein of interest or nucleic acids modulating host gene expression are expressed under control of the arenavirus promoters, internal ribosome entry sites or under control of regulatory elements that can be read by the viral RNA-dependent RNA polymerase, cellular RNA polymerase I, RNA polymerase II or RNA polymerase III. The modified arenaviruses are useful as vaccines and therapeutic agents for a variety of diseases.
1. A plasmid for expressing an arenavirus genome segment comprising a nucleic acid sequence that can express an arenavirus genome segment, wherein the arenavirus genome segment is an S segment or an L segment, and wherein an arenavirus open reading frame of the arenavirus genome segment that encodes a glycoprotein (GP), a nucleoprotein (NP), a matrix protein Z or a RNA-dependent RNA polymerase is removed or functionally inactivated.
2. The plasmid of claim 1 , wherein the arenavirus genome segment is engineered to have the ability to amplify and express its genetic information in arenavirus infected cells.
3. The plasmid of claim 1 , wherein the arenavirus genome segment is an S segment and the arenavirus open reading frame of the arenavirus genome segment that encodes the GP is removed or functionally inactivated.
4. The plasmid of claim 3 , wherein the arenavirus open reading frame encoding the GP is removed and replaced with a foreign ribonucleic acid sequence encoding a protein or peptide of interest.
5. A complementing cell for the production of an arenavirus particle, wherein the complementing cell comprises a plasmid of claim 1 , and expresses the open reading frame that has been removed or functionally inactivated from the arenavirus genome.
6. A method for producing replication-deficient infectious arenavirus particles comprising culturing the complementing cell of claim 5 and harvesting the culture supernatant comprising said arenavirus particles.