Alphavirus replicons and helper constructs

A recombinant replicon nucleic acid comprising:

• a) a nucleic acid sequence encoding a 5′ alphavirus replication recognition sequence;
• b) a nucleic acid sequence encoding an alphavirus nonstructural protein;
• c) an alphavirus subgenomic promoter-IRES-heterologous nucleic acid of interest (NOI) cassette, said cassette further comprising a spacer non-coding nucleic acid 3′ to the alphavirus subgenomic promoter and 5′ to the IRES;
• and
• d) a nucleic acid encoding a 3′ alphavirus replication recognition sequence.

A recombinant nucleic acid comprising:

• a) a 5′ alphavirus replication recognition sequence;
• b) an alphavirus subgenomic promoter-IRES-heterologous NOI cassette, which is in the 5′ to 3′ orientation, wherein the NOI encodes one or more alphavirus structural proteins;
• and
• c) a 3′ alphavirus replication recognition sequence.

A recombinant replicon nucleic acid comprising:

• a) a nucleic acid sequence encoding a 5′ alphavirus replication recognition sequence;
• b) a nucleic acid sequence encoding an alphavirus nonstructural protein;
• c) a first alphavirus subgenomic promoter-IRES-heterologous NOI cassette, which is in the 5′ to 3′ orientation;
• d) a second alphavirus subgenomic promoter-IRES-heterologous NOI cassette, which is in the 5′ to 3′ orientation;
• and
• e) a nucleic acid encoding a 3′ alphavirus replication recognition sequence.

A recombinant replicon nucleic acid comprising:

• a) a nucleic acid sequence encoding a 5′ alphavirus replication recognition sequence;
• b) a nucleic acid sequence encoding an alphavirus nonstructural protein;
• c) a first alphavirus subgenomic promoter-IRES-heterologous NOI cassette, said cassette further comprising a first spacer non-coding nucleic acid 3′ to the alphavirus subgenomic promoter and 5′ to the IRES;
• d) a second alphavirus subgenomic promoter-IRES-heterologous NOI cassette, said cassette further comprising a first spacer non-coding nucleic acid 3′ to the alphavirus subgenomic promoter and 5′ to the IRES;
• and
• e) a nucleic acid encoding a 3′ alphavirus replication recognition sequence.