Journal
NATURE BIOTECHNOLOGY
Volume 18, Issue 5, Pages 527-532Publisher
NATURE AMERICA INC
DOI: 10.1038/75390
Keywords
adeno-associated virus vector; gene therapy; hepatocytes; intermolecular recombination; concatemer
Categories
Funding
- NHLBI NIH HHS [R0I HL53682] Funding Source: Medline
Ask authors/readers for more resources
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 1 alpha (EF1 alpha) gene enhancer/promoter(s) (EF1 alpha EP) with a splice donor. We co-injected a promoter-less nlslacZ vector with a vector containing either a single EF1 alpha EP or a double copy of the EF1 alpha EP 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 EF1 alpha EP/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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available