Back to results
US 7,034,141 B2Granted
Packaging of positive-strand RNA virus replicon particles
AssigneeWyeth Holdings Corporation
InventorsGerald R. Kovacs, Nikos Vasilakis, Jacek Kowalski, Seema Gangolli, Timothy Zamb
Life of this patent
Estimating the term…
Abstract
The invention generally relates to recombinant polynucleotides, positive-strand RNA virus (psRNAV) recombinant expression vectors, and packaging systems. The packaging systems are based on the expression of helper functions by coinfecting recombinant poxvirus vectors comprising recombinant polynucleotides. Methods for obtaining psRNAV replicon particles using these packaging systems are disclosed. Immunogenic compositions and pharmaceutical formulations are provided that comprise replicon particles of the invention. Methods for generating an immune response or producing a pharmaceutical effect are also provided.
Claims37
A recombinant DNA polynucleotide comprising:
- a first portion comprising a DNA sequence encoding a DNA-dependent RNA polymerase operatively linked to a first heterologous promoter;
- and a second portion comprising either (i) a sequence encoding at least one structural protein of a positive-strand RNA virus (psRNAV), but not all of the structural proteins of said psRNAV, operatively linked to a second heterologous promoter or (ii) a second portion comprising a DNA sequence encoding a replicon-like psRNAV helper RNA sequence operatively linked to a second heterologous promoter;
- or (iii) a second portion comprising a DNA sequence encoding a replicon-like PSRNAV helper RNA sequence comprising a reporter gene operatively linked to a second heterologous promoter.
A recombinant DNA polynucleotide comprising:
- a first portion comprising a sequence encoding at least one structural protein of a psRNAV, but not all of the structural proteins of said psRNAV, operatively linked to a first heterologous promoter;
- and a second portion comprising a second heterologous promoter operatively linked to a psRNAV replicon comprising an psRNAV subgenomic promoter operatively linked to a sequence encoding at least one foreign polypeptide.
A recombinant DNA polynucleotide comprising:
- a first portion comprising a DNA sequence encoding a replicon-like psRNAV helper RNA sequence operatively linked to a first heterologous promoter;
- and a second portion comprising a second heterologous promoter operatively linked to a psRNAV replicon comprising a psRNAV subgenomic promoter operatively linked to a sequence encoding at least one foreign polypeptide.
A psRNAV replicon packaging system comprising:
- (a) a recombinant MVA comprising a recombinant polynucleotide comprising:
- a first portion comprising a sequence encoding a DNA-dependent RNA polymerase operatively linked to a first heterologous promoter;
- and a second portion comprising a sequence encoding at least one structural protein of a positive-strand RNA virus (psRNAV), but not all of the structural proteins of said psRNAV, operatively linked to a second heterologous promoter, wherein the at least one psRNAV structural protein of the second portion is selected from a psRNAV capsid and a psRNAV glycoprotein;
- and
- (b) a recombinant MVA comprising a recombinant polynucleotide comprising:
- a first portion comprising a sequence encoding at least one structural protein of a psRNAV, but not all of the structural proteins of said psRNAV, operatively linked to a first heterologous promoter;
- and a second portion comprising a second heterologous promoter operatively linked to a psRNAV replicon comprising an psRNAV subgenomic promoter operatively linked to a sequence encoding at least one foreign polypeptide;
- wherein the at least one psRNAV structural protein of the first portion is selected from a psRNAV capsid and a psRNAV glycoprotein;
- wherein the psRNAV structural protein of (a) and the psRNAV structural protein of (b) are not the same.
6 more elements
A psRNAV replicon packaging system comprising:
- (a) a recombinant MVA comprising a recombinant polynucleotide comprising:
- a first portion comprising a sequence encoding a DNA-dependent RNA polymerase operatively linked to a first heterologous promoter;
- and a second portion comprising a sequence encoding a replicon-like psRNAV helper RNA sequence operatively linked to a second heterologous promoter;
- wherein the replicon-like psRNAV helper RNA sequence of the second portion is a sequence encoding a psRNAV structural protein selected from a psRNAV capsid and a psRNAV glycoprotein;
- and
- (b) a recombinant MVA comprising a recombinant polynucleotide comprising:
- a first portion comprising a sequence encoding a replicon-like psRNAV helper RNA sequence operatively linked to a first heterologous promoter;
- and a second portion comprising a second heterologous promoter operatively linked to a psRNAV replicon comprising a psRNAV subgenomic promoter operatively linked to a sequence encoding at least one foreign polypeptide, wherein the replicon-like psRNAV helper RNA sequence of the first portion is a sequence encoding a psRNAV structural protein selected from a psRNAV capsid and a psRNAV glycoprotein;
- wherein the psRNAV structural protein of (a) and the psRNAV structural protein of (b) are not the same.
6 more elements
A psRNAV replicon packaging system comprising:
- (a) a first recombinant vector comprising a recombinant DNA polynucleotide comprising:
- a first portion comprising a sequence encoding a DNA-dependent RNA polymerase operatively linked to a first heterologous promoter;
- and a second portion comprising either (i) a sequence encoding at least one structural protein of a positive-strand RNA virus (psRNAV), but not all of the structural proteins of said psRNAV, operatively linked to a second heterologous promoter or (ii) a second portion comprising a DNA sequence encoding a replicon-like psRNAV helper RNA sequence operatively linked to a second heterologous promoter, wherein the at least one psRNAV structural protein of the second portion is selected from a psRNAV capsid and a psRNAV glycoprotein;
- and
- (b) a second recombinant vector comprising a recombinant DNA polynucleotide comprising:
- a first portion comprising a sequence encoding at least one structural protein of a psRNAV, but not all of the structural proteins of said psRNAV, operatively linked to a first heterologous promoter;
- and a second portion comprising a second heterologous promoter operatively linked to a psRNAV replicon comprising an psRNAV subgenomic promoter operatively linked to a sequence encoding at least one foreign polypeptide, wherein the at least one psRNAV structural protein of the first portion is selected from a psRNAV capsid and a psRNAV glycoprotein;
- wherein the psRNAV structural protein of (a) and the psRNAV structural protein of (b) are not the same;
- and wherein the first recombinant vector and the second recombinant vector are not derived from the same plasmid or virus vector.
6 more elements
A recombinant DNA polymycleotide comprising:
- a first portion comprising a first heterologous promoter operatively linked to a first sequence encoding a psRNAV capsid;
- and a second portion comprising a second heterologous promoter operatively linked to a second sequence encoding a psRNAV glycoprotein.
Dependencies are inferred from the claim text.