Gene transfer vector, recombinant adenovirus particles containing the same, method for producing the same and method of use of the same
First Claim
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1. A gene transfer vector consisting essentially of, in 5'"'"' to 3'"'"' orientation, the following elements:
- (i) a first adenovirus inverted terminal repeat,(ii) an adenoviral VAI gene and/or VAII gene,(iii) a gene foreign to adenovirus, wherein said gene is operably linked to a promoter functional in adenovirus target cells, and(iv) a second adenovirus inverted terminal repeat,wherein the order of elements (ii) and (iii) may be reversed; and
wherein one or both of element (i) and element (iv) additionally comprise an adenovirus packaging signal, and wherein said vector is incapable of producing, in vitro, recombinant adenovirus virus particles which have encapsidated therein said vector unless said vector is co-transfected or co-infected into adenovirus host cells with adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively.
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Abstract
A gene transfer vector comprising adenovirus inverted terminal repeats, at least one adenovirus packaging signal, and an adenoviral VAI gene and/or VAII gene; recombinant adenovirus particles containing the same; a method for producing the same and a method of use of the same to introduce and express a foreign gene in adenovirus target cells, is disclosed.
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Citations
36 Claims
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1. A gene transfer vector consisting essentially of, in 5'"'"' to 3'"'"' orientation, the following elements:
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(i) a first adenovirus inverted terminal repeat, (ii) an adenoviral VAI gene and/or VAII gene, (iii) a gene foreign to adenovirus, wherein said gene is operably linked to a promoter functional in adenovirus target cells, and (iv) a second adenovirus inverted terminal repeat, wherein the order of elements (ii) and (iii) may be reversed; and
wherein one or both of element (i) and element (iv) additionally comprise an adenovirus packaging signal, and wherein said vector is incapable of producing, in vitro, recombinant adenovirus virus particles which have encapsidated therein said vector unless said vector is co-transfected or co-infected into adenovirus host cells with adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A recombinant adenovirus virus particle which has encapsidated therein a gene transfer vector consisting essentially of, in 5'"'"' to 3'"'"' orientation, the following elements:
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(i) a first adenovirus inverted terminal repeat, (ii) an adenoviral VAI gene and/or VAII gene, (iii) a gene foreign to adenovirus, wherein said gene is operably linked to a promoter functional in adenovirus target cells, and (iv) a second adenovirus inverted terminal repeat, wherein the order of elements (ii) and (iii) may be reversed; and
wherein one or both of element (i) and element (iv) additionally comprise an adenovirus packaging signal, and wherein said vector is incapable of producing, in vitro, recombinant adenovirus virus particles which have encapsidated therein said vector unless said vector is co-transfected or co-infected into adenovirus host cells with adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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17. A method for the production of recombinant adenovirus particles comprising the steps of:
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(1) co-transfecting or co-infecting adenovirus host cells with (A) adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively, wherein said genomic DNA encodes a defective adenovirus VAI gene and/or VAII gene, and (B) a gene transfer vector consisting essentially of, in 5'"'"' to 3'"'"' orientation, the following elements; (i) a first adenovirus inverted terminal repeat, (ii) an adenoviral VAI gene and/or VAII gene, (iii) a gene foreign to adenovirus, wherein said gene is operably linked to a promoter functional in adenovirus target cells, and (iv) a second adenovirus inverted terminal repeat, wherein the order of elements (ii) and (iii) may be reversed; and
wherein one or both of element (i) and element (iv) additionally comprise an adenovirus packaging signal, and wherein said vector is incapable of producing, in vitro, recombinant adenovirus virus particles which have encapsidated therein said vector unless said vector is co-transfected or co-infected into adenovirus host cells with adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively;(2) harvesting adenovirus particles produced by the resulting host cells; and (3) separating, by CsCl centrifugation, adenovirus particles which have encapsidated therein said adenovirus genomic DNA from recombinant adenovirus particles which have encapsidated therein said gene transfer vector. - View Dependent Claims (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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29. A method for introducing and expressing a foreign gene in adenovirus target cells comprising infecting said adenovirus target cells with a recombinant adenovirus particle which has encapsidated therein a gene transfer vector consisting essentially of, in 5'"'"' to 3'"'"' orientation, the following elements:
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(i) a first adenovirus inverted terminal repeat, (ii) an adenoviral VAI gene and/or VAII gene, (iii) a gene foreign to adenovirus, wherein said gene is operably linked to a promoter functional in adenovirus target cells, and (iv) a second adenovirus inverted terminal repeat, wherein the order of elements (ii) and (iii) may be reversed; and
wherein one or both of element (i) and element (iv) additionally comprise an adenovirus packaging signal, and wherein said vector is incapable of producing, in vitro, recombinant adenovirus virus particles which have encapsidated therein said vector unless said vector is co-transfected or co-infected into adenovirus host cells with adenovirus genomic DNA or adenovirus particles containing adenovirus genomic DNA, respectively. - View Dependent Claims (30, 31, 32, 33, 34, 35, 36)
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Specification