Retroviral vector for the administration and expression of replicon rna expressing heterologous nucleic acids
AssigneeImmune Design Corp.
InventorsPeter Lars Aksel Berglund, Jacob Freeman Archer, Tsai-yu Lin
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Abstract
The present disclosure relates generally to gene delivery using a chimeric, retroviral-RNA replicon vector particle for increased expression of transgenes in a host cell. In particular, the chimeric vectors described herein can be used in any of a variety of settings including gene therapy and vaccine settings.
Claims31
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A chimeric retroviral-RNA replicon vector particle, comprising:
- a) an envelope glycoprotein;
- b) retroviral gag polyprotein comprising matrix, capsid and nucleocapsid proteins;
- c) a retroviral protease;
- and
- d) an alphaviral replicon sequence comprising in a 5′ to 3′ direction:
- i) a polynucleotide sequence comprising an alphavirus replication signal comprising at least a 5′ untranslated region (5′UTR) and a first copy of a polynucleotide sequence comprising a portion of the nsP1 coding sequence comprising at least a conserved sequence element (CSE);
- ii) a retroviral packaging sequence;
- iii) an IRES;
- iv) a polynucleotide sequence encoding a full-length replicase polyprotein (REP) comprising a portion of the nsP1 coding sequence comprising at least the CSE, wherein the second copy of the polynucleotide sequence is codon modified and in frame with the remainder of the sequence encoding the full-length REP;
- and v) a heterologous nucleic acid sequence of interest (SOI), wherein the SOI does not encode a), b), or c).
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A packaging system for producing a chimeric retroviral-RNA replicon vector particle, wherein the particle is reverse transcriptase independent, comprising a packaging cell transfected or otherwise modified to contain:
- a) a first nucleic acid molecule encoding an envelope;
- b) a second nucleic acid molecule encoding gag and pol proteins, wherein the nucleic acid molecule optionally encodes a nonfunctional reverse transcriptase protein;
- c) a third nucleic acid molecule comprising an alphaviral replicon sequence comprising in a 5′ to 3′ direction:
- i) a polynucleotide sequence comprising an alphavirus replication signal comprising at least a 5′ untranslated region (5′UTR) and a first copy of a polynucleotide sequence comprising a portion of the nsP1 coding sequence which comprises at least a conserved sequence element (CSE);
- ii) a retroviral packaging sequence;
- iii) an IRES;
- iv) a polynucleotide sequence encoding a full-length replicase polyprotein (REP) comprising a portion of the nsP1 coding sequence comprising at least the CSE, wherein the second copy of the polynucleotide sequence is codon modified and in frame with the remainder of the sequence encoding the full-length REP;
- and v) a heterologous nucleic acid sequence of interest (SOI).
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Dependencies are inferred from the claim text.