Patent ID: 6080569
Filing Date: 2000-06-27
Classification: A01K,C12N

Abstract:
A method for producing packaged, high-cloning capacity adenovirus virions which comprises:(a) introducing into a mammalian host cell which does not encode or contain active wild-type pIX protein, such that said cell is permissive for packaging of a helper-dependent adenovirus vector into and replication and production of said adenovirus virions, but non-permissive for packaging of DNA of a helper adenovirus:(i) a packaging-defective helper adenovirus comprising: a genome size greater than about 35 kb, sufficient adenovirus DNA to allow replication of a helper-dependent adenovirus vector, and a deletion or mutation of adenovirus DNA encoding pIX, wherein the DNA of said helper adenovirus is incapable of being packaged in the absence of wild type pIX protein, and(ii) a helper-dependent adenovirus vector incapable of viral replication in the absence of virion proteins encoded by said helper virus, wherein said vector comprises: a genome size less than about 35 kb, an adenoviral left ITR, an adenoviral right ITR, an adenoviral cis-acting packaging signal, no active or wild-type pIX encoding sequences, and at least one foreign DNA insert, and further wherein said vector is capable of being packaged into said virions under conditions in which the DNA of said helper adenovirus is not packaged; and(b) recovering said virions;wherein said packaging signal in said helper adenovirus is flanked by loxP sites recognizable by Cre recombinase.