Journal article
Expanding AAV Packaging Capacity with Trans-splicing or Overlapping Vectors: A Quantitative Comparison
Molecular therapy, Vol.4(4), pp.383-391
10/2001
DOI: 10.1006/mthe.2001.0456
PMID: 11592843
Abstract
Recombinant adeno-associated (rAAV) viral vectors hold great therapeutic potential for human diseases. However, a relatively small packaging capacity (less than 5 kb) has limited the application of rAAV for certain diseases such as cystic fibrosis and Duchenne muscular dystrophy. Here we compared two mechanistically distinct approaches to overcome packaging restraints with rAAV vectors. The trans-splicing approach reconstitutes gene expression from two independent rAAV vectors, each encoding unique, nonoverlapping halves of a transgene. This process requires intermolecular concatamerization and subsequent splicing between independent vectors. A distinct overlapping vector approach uses homologous recombination between overlapping regions in two independent vectors. Using the β-galactosidase gene as template, trans-splicing approaches were threefold (in primary fibroblasts) and 12-fold (in muscle tissue) more effective in generating full-length transgene products than the overlapping vector approach. Nevertheless, the efficiency of trans-splicing remained moderate at approximately 4.3% (for muscle) and 7% (for fibroblasts) of that seen with a single vector encoding the full-length transgene. The efficiency of trans-splicing was augmented 1185-fold by adenoviral E4, but not E2a, gene products. This augmentation was much less pronounced with the overlapping vectoring approach (12-fold). Trans-splicing and overlapping vector approaches are two viable alternatives to expand rAAV packaging capacity.
Details
- Title: Subtitle
- Expanding AAV Packaging Capacity with Trans-splicing or Overlapping Vectors: A Quantitative Comparison
- Creators
- Dongsheng Duan - Department of Anatomy & Cell Biology, The University of Iowa, Iowa City, Iowa, 52242, USAYongping Yue - Department of Anatomy & Cell Biology, The University of Iowa, Iowa City, Iowa, 52242, USAJohn F Engelhardt - Department of Anatomy & Cell Biology, The University of Iowa, Iowa City, Iowa, 52242, USA
- Resource Type
- Journal article
- Publication Details
- Molecular therapy, Vol.4(4), pp.383-391
- Publisher
- Elsevier Inc
- DOI
- 10.1006/mthe.2001.0456
- PMID
- 11592843
- ISSN
- 1525-0016
- eISSN
- 1525-0024
- Language
- English
- Date published
- 10/2001
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984025371302771
Metrics
20 Record Views