Method and means for producing high titer, safe, recombinant lentivirus vectors
First Claim
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1. A human immunodeficiency virus (HIV) transfer vector comprising a 5'"'"' LTR and a 3'"'"' LTR, each of which contains a U3 region, wherein a part or all of a regulatory element of the U3 region of the 5'"'"' LTR is replaced by another regulatory element, operable in a mammalian cell, which is not endogenous to said HIV.
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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.
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16 Claims
- 1. A human immunodeficiency virus (HIV) transfer vector comprising a 5'"'"' LTR and a 3'"'"' LTR, each of which contains a U3 region, wherein a part or all of a regulatory element of the U3 region of the 5'"'"' LTR is replaced by another regulatory element, operable in a mammalian cell, which is not endogenous to said HIV.
- 9. A human immunodeficiency virus (HIV) packaging plasmid lacking sequences upstream from gag endogenous to said HIV and lacking sequences downstream from env endogenous to said HIV.
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13. A method for producing a recombinant human immunodeficiency virus (HIV) vector comprising:
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a) transforming a cell with; i) at least one HIV packaging plasmid lacking sequences upstream from gag endogenous to said HIV and lacking sequences downstream from env endogenous to said HIV, and said at least one packaging plasmid which comprises a gag, a pol or gag and pol genes; and ii) an expression plasmid not endogenous to said HIV which carries an env gene not endogenous to said HIV; to yield a packaging cell; b) multiply transforming said packaging cell with an HIV 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 HIV vector from said medium. - View Dependent Claims (14, 15, 16)
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Specification