Engineered adenovirus vectors and uses thereof
OwnerRichard Lowenthalinventor
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Abstract
Disclosed herein relates to a replication competent recombinant adenovirus comprising restriction enzyme sites in an E3 region to insert a gene of interest. Also disclosed herein is a method for generating the replication competent recombinant adenovirus and a pharmaceutical composition comprising the adenovirus for use in treating a disease or a condition.
Claims65
A replication competent recombinant adenovirus comprising an I-CeuI restriction site, a PI-SceI restriction site, or a combination thereof in an E3 region.
The adenovirus of any one of the preceding claims, wherein the adenovirus is selected from the group consisting of adenovirus type 2, adenovirus type 4, adenovirus type 5, and adenovirus type 7.
The adenovirus of any one of the preceding claims, wherein the adenovirus preferably replicates in a tumor cell as compared to a non-tumor cell.
The adenovirus of any one of the preceding claims comprising an I-CeuI restriction site and a PI-SceI restriction site in the E3 region.
A pharmaceutical composition comprising the adenovirus of any one of the preceding claims and a pharmaceutically acceptable carrier.
A method for generating a replication competent recombinant adenovirus comprising a heterologous nucleotide sequence, comprising:
- (a) cloning a heterologous nucleotide sequence into a shuttle vector;
- (b) digesting the shuttle vector in (a) with an I-CeuI restriction enzyme and a PI-SceI restriction enzyme to excise a fragment encompassing the heterologous nucleotide sequence;
- (c) digesting an adenovirus vector with the I-CeuI restriction enzyme and the PI-SceI restriction enzyme;
- (d) ligating the fragment encompassing the heterologous nucleotide sequence in (b) into the adenovirus vector in (c), thereby generating a recombinant adenoviral plasmid;
- (e) transfecting the recombinant adenoviral plasmid into a packaging cell;
- and
- (f) recovering the replication competent recombinant adenovirus from the packaging cell, wherein the adenovirus vector comprises an I-CeuI restriction site and a PI-SceI restriction site in an E3 region.
A polynucleotide sequence comprising an E3 region of an adenovirus, wherein the E3 region comprises an I-CeuI restriction site, a PI-SceI restriction site, or a combination thereof.
A modified E3 region of an adenovirus, wherein the modified E3 region comprises an I-CeuI restriction site, a PI-SceI restriction site, or a combination thereof.
Dependencies are inferred from the claim text.