Patent ID: 6566128
Filing Date: 2003-05-20
Classification: A01K,C12N

Abstract:
A method for producing packaged, high-cloning capacity adenovirus virions, wherein said method comprises:(A) introducing into a mammalian host cell, wherein said mammalian host cell does not encode or contain active or wild-type pIX protein and wherein said mammalian host cell is permissive for packaging DNA of a helper-dependent adenovirus vector into and replication and production of said adenovirus virions, but non-permissive for packaging of DNA of a pIX-defective helper adenovirus: (i) said DNA of said pIX-defective helper adenovirus, wherein said DNA comprises: a. a size greater than about 35 kb; and b. a deletion or mutation in the DNA sequence encoding pIX, wherein the DNA of said pIX-defective helper adenovirus is unpackageable in the absence of active or wild type pIX protein, and (ii) said DNA of said helper-dependent adenovirus vector, wherein said DNA of said helper-dependent vector comprises: a a size less than about 35 kb; b an adenoviral left ITR; c an adenoviral right ITR; d an adenoviral cis-acting packaging signal; e no active or wild-type sequences encoding pIX; and f at least one foreign DNA insert, wherein said DNA of said helper-dependent adenovirus vector replicates, and is packaged into said adenovirus virions; and (B) recovering said adenovirus virions, wherein said adenovirus virions have a genome size at least about 15 kb and at most about 35 kb.