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US 2003/0148522 A1Published application
Lentivirus-based gene transfer vectors
OwnerJohn C. Olseninventor
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OwnerJohn C. Olseninventor
Estimating the term…
A recombinant lentiviral vector expression system comprises a first vector that comprises a nucleic acid sequence of at least part of the Equine Infectious Anemia Virus (EIAV) genome. The vector contains at least one defect in at least one gene encoding an EIAV structural protein, but is preferably a gag/pol expression vector. The expression system further comprises a second vector, also comprising a nucleic acid sequence of at least part of the Equine Infectious Anemia Virus (EIAV) genome, and additionally containing a multiple cloning site wherein a heterologous gene may be inserted. The expression system also comprises a third vector which expresses a viral envelope protein. The first and third vectors are packaging signal-defective. When the expression system is transfected into a lentivirus-permissive cell, replication-defective EIAV particles may be produced, which particles are useful in delivering heterologous genes to a broad range of both dividing and non-dividing cells.
A recombinant lentiviral vector expression system comprising:
The plasmid set forth in FIG. 2A as pEV53.
The plasmid set forth in FIG. 2B as pEV53A.
The plasmid set forth-in FIG. 2 as pEC lacZ.
The plasmid set forth in FIG. 3 as pEC-puro.
The plasmid set forth in FIG. 4 as pCI-VSV-G.
A method of producing a replication-defective lentivirus particle, comprising transfecting a cell with:
An infectious EIAV particle containing an EIAV nucleic acid sequence encoding a promoter and a heterologous gene sequence, and wherein said nucleic acid sequence is defective in encoding at least one EIAV structural protein so that said virus particle is replication defective.
A method of delivering a heterologous gene to a target cell, comprising transfecting said target cell with:
A method of producing a lentiviral stock comprising:
A method of making a packaging cell, comprising transfecting a lentivirus-permissive cell with a vector comprising a nucleic acid sequence of at least part of the Equine Infectious Anemia Virus (EIAV) genome, wherein said vector contains a defective packaging signal.
A packaging cell comprising a lentivirus-permissive host cell containing an EIAV nucleic acid sequence encoding at least one EIAV structural protein, wherein said nucleic acid sequence is packaging-signal defective, such that the cell itself is capable of producing at least one EIAV structural protein, but not capable of producing replication-competent infectious virus.
Dependencies are inferred from the claim text.