High-efficiency promoter-dependent transduction by adeno-associated virus type 6 vectors in mouse lung.

Publication Type:

Journal Article

Source:

Human gene therapy, Volume 18, Issue 4, p.344-54 (2007)

Keywords:

Alkaline Phosphatase, Alternative Splicing, Animals, CYTOMEGALOVIRUS, Dependovirus, Genetic Vectors, Globins, Humans, lung, MICE, Promoter Regions, Genetic, TISSUE DISTRIBUTION, Transduction, Genetic

Abstract:

The transduction efficiency of adeno-associated virus (AAV) vectors in various somatic tissues has been shown to depend heavily on the AAV type from which the vector capsid proteins are derived. Among the AAV types studied, AAV6 efficiently transduces cells of the airway epithelium, making it a good candidate for the treatment of lung diseases such as cystic fibrosis. Here we have evaluated the effects of various promoter sequences on transduction rates and gene expression levels in the lung. Of the strong viral promoters examined, the Rous sarcoma virus (RSV) promoter performed significantly better than a human cytomegalovirus (CMV) promoter in the airway epithelium. However, a hybrid promoter consisting of a CMV enhancer, beta-actin promoter and splice donor, and a beta-globin splice acceptor (CAG promoter) exhibited even higher expression than either of the strong viral promoters alone, showing a 38-fold increase in protein expression over the RSV promoter. In addition, we show that vectors containing either the RSV or CAG promoter expressed well in the nasal and tracheal epithelium. Transduction rates in the 90% range were achieved in many airways with the CAG promoter, showing that with the proper AAV capsid proteins and promoter sequences, efficient transduction can be achieved.