Stocks of recombinant, replication-deficient adenovirus free of replication-competent adenovirus
First Claim
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1. A replication-competent adenovirus-free stock of a recombinant adenoviral vector, wherein said adenoviral vector is deficient in one or more essential gene functions of one or more regions of the adenoviral genome selected from the group consisting of the E1, E2A , and E4 regions of the adenoviral genome.
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Abstract
The present invention provides multiply deficient adenoviral vectors and complementing cell lines. Also provided are recombinants of the multiply deficient adenoviral vectors and a therapeutic method, particularly relating to gene therapy, vaccination, and the like, involving the use of such recombinants.
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Citations
24 Claims
- 1. A replication-competent adenovirus-free stock of a recombinant adenoviral vector, wherein said adenoviral vector is deficient in one or more essential gene functions of one or more regions of the adenoviral genome selected from the group consisting of the E1, E2A , and E4 regions of the adenoviral genome.
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21. A system comprising (i) a cell having a cellular genome comprising at least one DNA sequence, which is operably linked to an inducible promoter or a repressible promoter, and which complements in trans for a deficiency in one or more essential gene functions of the E1 and E2A regions of an adenoviral genome, and (ii) an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 and E2A regions of said adenoviral genome, wherein there is insufficient overlap between said cellular genorne and said adenoviral genome to mediate a recombination event sufficient to result in a replication-competent adenovirus.
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22. A system comprising (i) a cell having a cellular genome comprising at least one DNA sequence, which is operably linked to an inducible promoter or a repressible promoter, and which complements in trans for a deficiency in one or more essential gene functions of the E1, E2A and E4 regions of an adenoviral genome, and (ii) an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1, E2A and E4 regions of said adenoviral genome, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event sufficient to result in a replication-competent adenovirus.
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23. A method of propagating an adenoviral vector that is deficient in one or more essential gene functions of one or more regions of an adenoviral genome, which method comprises propagating an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene functions of the E1 and E2A regions in a cell having a cellular genome comprising at least one DNA sequence that complements in trans said deficiency, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event resulting in a replication-competent adenoviral vector.
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24. A method of propagating an adenoviral vector that is deficient in one or more essential gene functions of one or more regions of an adenoviral genome, which method comprises propagating an adenoviral vector comprising an adenoviral genome having a deficiency in one or more essential gene finctions of the E1, E2A and E4 regions in a cell having a cellular genome comprising at least one DNA sequence that complements in trans said deficiency, wherein there is insufficient overlap between said cellular genome and said adenoviral genome to mediate a recombination event resulting in a replication-competent adenoviral vector.
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