Recombinant vesiculoviruses and their uses
AssigneeYale University
InventorJohn Rose
Life of this patent
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Abstract
The present invention provides recombinant replicable vesiculoviruses. The invention provides a method which, for the first time, successfully allows the production and recovery of replicable vesiculoviruses, as well as recombinant replicable vesiculoviruses, from cloned DNA, by a method comprising expression of the full-length positive-strand vesiculovirus antigenomic RNA in host cells. The recombinant vesiculoviruses do not cause serious pathology in humans, can be obtained in high titers, and have use as vaccines. The recombinant vesiculoviruses can also be inactivated for use as killed vaccines.
Claims9
1-43. (canceled)
A kit comprising in one or more containers (a) a first recombinant nucleic acid that can be transcribed in a host cell to produce an RNA molecule comprising a vesiculovirus antigenomic (+) RNA, in which a foreign RNA sequence is inserted into or replaces a region of the RNA nonessential for replication of the vesiculovirus (b) a second recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus N protein;
- (c) a third recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus L protein;
- and (d) a fourth recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus P protein.
A kit comprising in one or more containers (a) a first recombinant nucleic acid that can be transcribed in a host cell to produce an RNA molecule comprising a vesiculovirus antigenomic (+) RNA;
- (b) a second recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus N protein;
- (c) a third recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus L protein;
- and (d) a fourth recombinant nucleic acid encoding and capable of expressing in the host cell a vesiculovirus P protein.
A kit comprising in one or more containers (a) a recombinant nucleic acid that can be transcribed to produce an RNA molecule comprising a vesiculovirus antigenomic (+) RNA;
- and (b) a host cell that recombinantly expresses vesiculovirus N, P, and L proteins.
A kit comprising
- (a) in a first container a first DNA plasmid vector comprising the following operatively linked components:
- (i) a bacteriophage RNA polymerase promoter;
- (ii) a first DNA sequence that can be transcribed in a cell to produce an RNA molecule comprising (A) a vesiculovirus antigenomic (+) RNA in which a foreign RNA sequence is inserted into or replaces a region of the RNA nonessential for replication of the vesiculovirus, and (B) a ribozyme sequence immediately downstream of said antigenomic (+) RNA, that cleaves at the 3′ terminus of the antigenomic RNA;
- and
- (iii) a transcription termination signal for the RNA polymerase;
- (b) in a second container a second DNA plasmid vector comprising the following operatively linked components:
- (i) the bacteriophage RNA polymerase promoter;
- (ii) a second DNA sequence encoding an N protein of the vesiculovirus;
- and
- (iii) a second transcription termination signal for the RNA polymerase;
- (c) in a third container a third DNA plasmid vector comprising the following operatively linked components:
- (i) the bacteriophage RNA polymerase promoter;
- (ii) a third DNA sequence encoding a P protein of the vesiculovirus;
- and
- (iii) a third transcription termination signal for the RNA polymerase;
- (d) in a fourth container a fourth DNA plasmid vector comprising the following operatively linked components:
- (i) the bacteriophage RNA polymerase promoter;
- (ii) a fourth DNA sequence encoding an L protein of the vesiculovirus;
- and
- (iii) a fourth transcription termination signal for the RNA polymerase;
- and
- (e) in a fifth container a recombinant vaccinia virus comprising a sequence encoding the bacteriophage RNA polymerase.
51-74. (canceled)
Dependencies are inferred from the claim text.