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- Title
Expression of a Truncated Cystic Fibrosis Transmembrane Conductance Regulator with an AAV5-pseudotyped Vector in Primates.
- Authors
Fischer, Anne C.; Smith, Carolina I.; Cebotaru, Liudmila; Xuemei Zhang; Askin, Frederic B.; Wright, Jerry; Guggino, Sandra E.; Adams, Robert J.; Flotte, Terence; Guggino, William B.
- Abstract
Gene therapy using recombinant adeno-associated virus (rAAV2) vectors for cystic fibrosis has shown gene transfer and remarkable safety, yet indeterminate expression. A new construct has been characterized with a powerful exogenous promoter, the cytomegalovirus enhancer/chicken β-actin promoter, driving a truncated CF transmembrane conductance regulator (CFTR), pseudotyped in an AAV5 viral coat. Our goal is to demonstrate that airway delivery of a pseudotyped rAAV5 vector results in gene transfer as well as expression in non-human primates. Aerosolized pseudotyped rAAV5-ΔCFTR or rAAV5-GFP (green fluorescent protein) genes were delivered to four and six lungs, respectively. The pseudotyped rAAV5 vector did result in GFP gene transfer (1.005×106 copies/μg DNA on average) and quantifiable gene expression. Microscopy confirmed protein expression in airway epithelium. Similarly, the vector also resulted in vector-specific CFTR DNA (1.24×105 copies/μg) and mRNA expression. Immunoprecipitation and 32P phosphoimaging were used to demonstrate CFTR protein expression, as qualitatively enhanced beyond the barely detectable endogenous expression in untreated animals. Based on these promising studies, this CFTR minigene construct is a therapeutic candidate.Molecular Therapy (2007) 15 4, 756–763. doi:10.1038/sj.mt.6300059
- Subjects
GENE therapy; CYSTIC fibrosis; GENETIC disorders; VIRAL genetics; GENETIC transformation
- Publication
Molecular Therapy, 2007, Vol 15, Issue 4, p756
- ISSN
1525-0016
- Publication type
Article
- DOI
10.1038/sj.mt.6300059