Abstract
A major shortcoming to the use of adeno-associated virus (rAAV) vectors is their limited packaging size. To overcome this hurdle, we split an expression cassette and cloned it into two separate vectors. The vectors contained either a nuclear localizing Escherichia coli lacZ transgene (nlslacZ) with a splice acceptor, or the human elongation factor 1α (EF1α) gene enhancer/promoter(s) (EF1αEP) with a splice donor. We co-injected a promoter-less nlslacZ vector with a vector containing either a single EF1αEP or a double copy of the EF1αEP in a head-to-head orientation, into the portal vein of mice. Gene expression, measured by both transduction efficiency and quantitation of the recombinant protein, was as much as 60-70% of that obtained from mice that received a single vector containing a complete EF1αEP/nlslacZ expression cassette. This two-vector approach may allow development of gene therapy strategies that will carry exogenous DNA sequences with large therapeutic cDNAs and/or regulatory elements.
Original language | English (US) |
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Pages (from-to) | 527-532 |
Number of pages | 6 |
Journal | Nature biotechnology |
Volume | 18 |
Issue number | 5 |
DOIs | |
State | Published - May 2000 |
Externally published | Yes |
Keywords
- Adeno-associated virus vector
- Concatemer
- Gene therapy
- Hepatocytes
- Intermolecular recombination
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
- Molecular Medicine
- Biomedical Engineering