USE OF THE PIX PROMOTER FOR GENE THERAPY APPLICATIONS

Abstract

The purpose of this research was to determine whether expression of a non-pIX coding sequence from the pIX promoter would be replication-responsive. A pIX expression cassette was generated in adenovirus genome plasmids; the plasm ids were transfected into cells to produce infectious virus. SEAP expression from the pIX promoter was evaluated in plasmid and virus contexts. Infections were performed under permissive and, what were thought to be, non-permissive conditions. SEAP activity and vector genome copy number were measured. SEAP activity per genome was calculated for replicated and non-replicated genomes. SEAP activity per genome did not vary more than 2.25 fold between replicated and non-replicated genomes. It was determined that expression of a non-pIX transgene from the pIX promoter is not necessarily replicationdependent. Additionally, replication of an El-deleted adenovirus in an E1-/E4-deficient growth environment (A549 cells) was detected. No examples for comparison were found in the literature.