Method for the preparation of a viral vector by inter-molecular homologous recombination
AssigneeTransgene S.A.
InventorsCécile Chartier, Eric Degryse
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Abstract
The present invention encompasses methods of preparing a recombinant viral vector, within a procaryotic cell, by intermolecular homologous recombination, methods of preparing an infectious viral particle containing the recombinant viral vector and pharmaceutical compositions comprising said vector or particle. The invention also encompasses the therapeutic use of said vector or particle, especially in human gene therapy.
Claims17
A method for preparing a recombinant adenoviral vector containing an exogenous DNA sequence comprising the steps of:
- (a) introducing into a prokaryotic cell:
- (i) a first DNA fragment comprising all or part of an adenoviral genome, including the encapsidation region and the 3′ and 5′ITRs and
- (ii) a second DNA fragment comprising said exogenous DNA sequence surrounded by flanking sequences A and B which are homologous to (i), and
- (b) culturing the prokaryotic cell obtained in (a) under suitable culture conditions to allow intermolecular homologous recombination and insertion of the exogenous DNA into the first DNA fragment to occur and recovering the resulting recombinant adenoviral vector;
- wherein the recombinant adenoviral vector comprises said adenoviral genome including the encapsidation region and the 3′ and 5′ITRs and lacking all or part of the E1, E2, E3, and/or E4 regions and further comprising the exogenous DNA sequence.
A method for preparing a recombinant adenoviral vector containing an exogenous DNA sequence comprising the steps of:
- (a) introducing into a prokaryotic cell:
- (i) a first DNA fragment comprising all or part of an adenoviral genome, and
- (ii) a second DNA fragment comprising said exogenous DNA sequence surrounded by flanking sequences A and B which are homologous to (i), wherein the encapsidation region, the 3′ and/or 5′ITRs are comprised in the first or in the second DNA fragment and
- (b) culturing the prokaryotic cell obtained in (a) under suitable culture conditions to allow homologous recombination and insertion of the exogenous DNA into the first DNA fragment to occur, and recovering the resulting recombinant adenoviral vector;
- wherein the recombinant adenoviral vector comprises said adenoviral genome including the encapsidation region and the 3′ and 5′ITRs and lacking all or part of the E1, E2, E3 and/or E4 regions, and further comprises the exogenous DNA sequence.
Dependencies are inferred from the claim text.