Alphavirus vector constructs

An alphavirus vector construct, comprising a promoter 5' of viral cDNA which initiates the synthesis of RNA from the viral cDNA by a process of in vitro transcription, followed by a 5' sequence which initiates transcription of alphavirus RNA, followed by a nucleotide sequence encoding alphavirus non-structural proteins, a viral junction region which has been modified such that viral transcription of a subgenomic fragment is reduced, an internal ribosome entry site or a sequence which promotes ribosome readthrough between adjacent reading frames, and an alphavirus RNA polymerase recognition sequence.

An alphavirus vector construct, comprising a promoter 5' of viral cDNA which initiates the synthesis of RNA from the viral cDNA by a process of in vitro transcription, followed by a 5' sequence which initiates transcription of alphavirus RNA, followed by a nucleotide sequence encoding alphavirus non-structural proteins, a First viral junction region which has been inactivated such that viral transcription of a subgenomic fragment is prevented, a second viral junction region which has been modified such that viral transcription of the subgenomic fragment is reduced, an internal ribosome entry site or a sequence which promotes ribosome readthrough between adjacent reading frames, and an alphavirus RNA polymerase recognition sequence.

An alphavirus cDNA vector construct comprising a promoter 5' of viral cDNA which initiates the synthesis of RNA from the viral EDNA within a cell, followed by a 5' sequence which initiates transcription of alphavirus RNA, followed by a nucleotide sequence encoding alphavirus non-structural proteins, a viral junction region consisting of (i) all active viral junction region, (ii) a viral junction region which has been modified such that viral transcription of a subgenomic fragment is reduced, and (iii) a viral junction region which has been inactivated such that viral transcription of a subgenomic fragment is prevented, and an alphavirus RNA polymerase recognition sequence.

An alphavirus cDNA vector construct comprising a promoter 5' of viral cDNA which initiates the synthesis of RNA from the viral cDNA within a cell, followed by a 5' sequence which initiates transcription of alphavirus RNA, a nucleotide sequence encoding alphavirus non-structural proteins, a first viral junction region which has been inactivated such that viral transcription of the subgenomic fragment is prevented, followed by a second viral junction region which has been modified such that viral transcription of the subgenomic fragment is reduced, and an alphavirus RNA polymerase recognition sequence.

A recombinant alphavirus particle which, upon introduction into at BHK cell, produces an infected cell which is viable at least 72 hours after infection.

A recombinant alphavirus particle which, upon introduction into a BHK cell, produces an infected cell which is viable at least 72 hours after infection, said particle also carrying a vector construct which directs the expression of at least one antigen or modified form thereof in target cells infected with the alphavirus particle, wherein said antigen or modified form thereof stimulates an immune response within an animal.

A packaging cell line which inducibly expresses alphavirus structural proteins, and which, upon introduction of an alphavirus vector construct, produces recombinant alphavirus particles.

A packaging cell line suitable for packaging and production of an alphavirus vector, wherein the packaging cell line comprises an expression cassette which directs the expression of VSV-G.

A producer cell line suitable for packaging and production of a recombinant alphavirus particle, wherein the producer cell line comprises an expression cassette which directs the expression of gag/pol, an expression cassette which directs the expression of env, and an alphavirus vector construct containing a retroviral packaging sequence.

A producer cell line suitable for packaging and production of a recombinant alphavirus particle, wherein the producer cell line comprises one or more expression cassettes which direct the expression of non-alphaviral structural proteins, and an alphavirus vector construct comprising a packaging sequence corresponding to a virus from which the non-alphaviral structural proteins are derived.