Patent ID: 8460913
Filing Date: 2013-06-11
Classification: A61K,A61P,C12N

Abstract:
1. A method of making an alphavirus replicon particle, comprising introducing one or more of an isolated RNA molecule, which RNA comprises: a) an alphavirus 5′ replication recognition sequence, wherein at least one initiation codon has been removed from the 5′ replication recognition sequence; b) a nucleotide sequence encoding an alphavirus structural protein; and c) an alphavirus 3′ replication recognition sequence, with the proviso that the RNA molecule does not contain a promoter that directs transcription of the nucleotide sequence of (b), and wherein the alphavirus 5′ and 3′ replication recognition sequences of (a) and (c) direct replication of the entire RNA molecule in the presence of alphavirus nonstructural proteins; into a cell, wherein the RNA molecule comprises: i) an alphavirus 5′ replication recognition sequence, wherein at least one initiation codon has been removed from the 5′ replication recognition sequence; ii) a nucleotide sequence encoding an alphavirus structural protein; and iii) an alphavirus 3′ replication recognition sequence, with the proviso that the RNA molecule does not contain a promoter that directs transcription of the nucleotide sequence of (ii), and wherein the alphavirus 5′ and 3′ replication recognition sequences of (i) and (iii) direct replication of the entire RNA molecule in the presence of nonstructural proteins, whereby the combination of RNA molecules encodes all alphavirus structural proteins necessary for production of an alphavirus replicon particle, along with an alphavirus replicon RNA, under conditions whereby alphavirus replicon particles are produced.