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US 6,428,953 B1Granted
Method and means for producing high titer, safe, recombinant lentivirus vectors
AssigneeCell Genesys, Inc.
InventorsLuigi Naldini, Thomas Dull, Deborah A. Farson, Rochelle Witt
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Abstract
Lentiviral vectors modified at the 5′ LTR or both the 5′ and 3′ LTR's are useful in the production of recombinant lentivirus vectors. Such vectors can be produced in the absence of a functional tat gene. Multiple transformation of the host cell with the vector carrying the transgene enhances virus production.
Claims9
A lentivirus packaging plasmid lacking sequences upstream from gag endogenous to said lentivirus and lacking sequences downstream from env endogenous to said lentivirus.
A method for producing a recombinant lentivirus vector comprising:
- a) transforming a cell with:
- i) at least one lentivirus packaging plasmid lacking sequences upstream from gag endogenous to said lentivirus and lacking sequences downstream from env endogenous to said lentivirus, and said at least one packaging plasmid comprises a gag, a pol or gag and pol genes;
- and
- ii) an expression plasmid not endogenous to said lentivirus which carries an env gene not endogenous to said lentivirus;
- to yield a packaging cell;
- b) multiply transforming said packaging cell with a lentivirus transfer vector which comprises a heterologous gene to yield a producer cell;
- c) culturing said producer cell in a medium;
- and
- d) separating said producer cell from said medium to recover said recombinant lentivirus vector from said medium.
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Dependencies are inferred from the claim text.