Adenoviral vector system comprising Cre-loxP recombination
AssigneeAdVec, Inc.
InventorsFrank L. Graham, Martina Anton, Michael A. Rudnicki
Life of this patent
Estimating the term…
Abstract
This invention provides a method for making adenovirus cloning vectors which contain a recombinase target site that is useful for the insertion of selected foreign proteins.
Claims6
An adenovirus vector system for expressing foreign DNA sequences, comprising an isolated cell co-infected with:
- a) a helper adenovirus comprising an E1 region where the packaging signals contained within the E1 region are flanked on both sides by lox P recombinase target sites;
- and
- b) a helper-dependent adenovirus vector comprising:
- 1) a deletion of up to approximately 35,000 bp of the adenovirus genome but retaining sufficient left and right ITR to support viral replication and packaging;
- and
- 2) a fragment or fragments of foreign DNA sequence of up to about 35,000 bp;
- and wherein the cell supports replication of the helper adenovirus, and where the cell additionally expresses Cre recombinase.
A method of making a helper adenovirus genome comprising infecting an isolated cell with an adenovirus comprising an E1 region where the packaging signals are flanked on both sides by lox P sites, wherein the cell expresses Cre recombinase, and incubating the infected cells such that the Cre recombinase excises the packaging signals such that the helper adenovirus genome does not package.
A method of making a packaged helper-dependent adenovirus vector comprising co-infecting an isolated cell with:
- a) a helper adenovirus vector comprising an E1 region where the packaging signals contained within the E1 region are flanked on both sides by lox P recombinase target sites;
- b) a helper-dependent adenovirus vector comprising:
- 1) a deletion of up to approximately 35,000 bp of the adenovirus genome but retaining an adenoviral packaging signal and sufficient left and right ITR to support viral replication and packaging;
- and
- 2) a fragment or fragments of foreign DNA sequence of about 35,000 bp, wherein the cell supports replication of the helper adenovirus, and
- where the cell expresses Cre recombinase, and incubating the co-infected cells such that the Cre recombinase catalyzes the removal of the packaging signals from the helper adenovirus vector such that it replicates but does not package, and wherein the helper virus supports replication of the helper-dependent adenovirus vector, and wherein the helper-dependent adenovirus vector is packaged into adenovirus virions.
Dependencies are inferred from the claim text.