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US 6,277,633 B1Granted
Lentivirus-based gene transfer vectors
AssigneeThe University of North Carolina at Chapel Hill
InventorJohn C. Olsen
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AssigneeThe University of North Carolina at Chapel Hill
InventorJohn C. Olsen
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 gaglpol 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 comprising a nucleic acid sequence encoding a promoter and a gene sequence heterologous to EIAV, 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 producing a lentiviral stock comprising:
A method of making a packaging cell, comprising transfecting a lentivirus-permissive cell with a vector comprising an EIAV nucleic acid sequence, wherein said vector comprises a defective packaging signal.
A packaging cell comprising a lentivirus-permissive host cell comprising 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 produces at least one EIAV structural protein, but does not produce replication-competent infectious virus.
Dependencies are inferred from the claim text.