Patent ID: 6168943
Filing Date: 2001-01-02
Classification: A61K,A61P,C07K,C12N

Abstract:
A method of producing a recombinant replicable vesiculovirus comprising culturing a cell containing (a) a first recombinant nucleic acid that can be transcribed to produce an RNA molecule comprising (A) a vesiculovirus antigenomic (+) RNA containing the vesiculovirus promoter for replication, and (B) a ribozyme sequence immediately downstream of said antigenomic (+) RNA, that cleaves at the 3' terminus of the antigenomic (+) RNA, in which a region of the RNA nonessential for replication of the vesiculovirus has been inserted into or replaced by a foreign RNA sequence, said first recombinant nucleic acid having the following operatively linked components: (i) a first promoter; (ii) a first DNA sequence that can be transcribed under the control of the first promoter in the cell to produce said RNA molecule; and (iii) a first transcription termination signal; (b) a second recombinant nucleic acid encoding a vesiculovirus N protein and having the following operatively linked components: (i) a second promoter which controls the expression of the N protein; (ii) a first translation initiation signal; and (iii) a second DNA sequence encoding the N protein; (c) a third recombinant nucleic acid encoding a vesiculovirus L protein and having the following operatively linked components: (i) a third promoter which controls the expression of the L protein; (ii) a second translation initiation signal; and (iii) a third DNA sequence encoding the L protein; (d) a fourth recombinant nucleic acid encoding a vesiculovirus P protein and having the following operatively linked components: (i) a fourth promoter which controls the expression of the P protein; (ii) a third translation initiation signal; and (iii) a fourth DNA sequence encoding the P protein; whereby the first recombinant nucleic acid is transcribed in the cell to produce said RNA molecule, and the N, L and P proteins are expressed in the cell, and a recombinant replicable vesiculovirus is produced that has a genome that is the complement of said antigenomic RNA comprising said foreign RNA sequence, in which said foreign RNA sequence encodes a peptide or protein that is expressed and induces an immune response to said peptide or protein in a suitable host infected by the vesiculovirus.