Methods of propagating monkey adenoviral vectors
View Patent ↗The invention provides methods for propagating a monkey adenovirus in a cell, including a human cell, comprising one or more gene products isolated from a human adenovirus. Also provided are methods for propagating wherein the monkey adenovirus comprises a nucleic acid sequence encoding a human adenovirus gene product. The invention further provides a monkey adenovirus, including a replication-deficient monkey adenovirus, obtained by such propagation methods.
1. A method of propagating a monkey adenovirus, which method comprises contacting a cell with the monkey adenovirus, wherein the monkey adenovirus comprises a nucleic acid sequence encoding one or more gene products of a human adenovirus,
wherein the one or more gene products comprise a gene product encoded by a portion of the E4 region responsible for alleviating or overcoming host replication block in human cells, which portion consists essentially of E4 ORF6, and whereby the monkey adenovirus is propagated in the cell,
wherein the monkey adenovirus further comprises inverted terminal repeat (ITR) sequences from the monkey adenovirus, wherein monkey refers to both new world and old world monkeys, and does not include any member of the family Hominidae.
2. The method of claim 1 , wherein the monkey adenovirus comprises a nucleic acid sequence encoding a species C human adenovirus gene product.
3. The method of claim 2 , wherein the one or more gene products comprise a gene product encoded by one or both of the E1A region and the E1B region of a human adenovirus.
4. The method of claim 3 , wherein the portion of the E4 region is E4 ORF6.
5. The method of claim 4 , wherein the cell is a human cell.
6. The method of claim 5 , wherein the cell is a HEK-293 cell or a PerC.6 cell.
7. The method of claim 1 , wherein the monkey adenovirus is replication-deficient.
8. The method of claim 7 , wherein the monkey adenovirus requires complementation of one or more of the E1A region, the E1B region, and the E4 region of the adenoviral genome for propagation.
9. The method of claim 8 , wherein the monkey adenovirus comprises a deficiency in the E1 region and a deficiency in at least a portion of the E4 region of the adenoviral genome.
10. The method of claim 9 , wherein the monkey adenovirus further comprises a deficiency in the E3 region of the adenoviral genome.