Efficient gene transfer into human keratinocytes with recombinant adeno-associated virus vectors
| Autor: | H Smola, M Braun-Falco, A Doenecke, Michael Hallek |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Keratinocytes
viruses Genetic enhancement Genetic Vectors Green Fluorescent Proteins Gene Expression Biology Transfection medicine.disease_cause Recombinant virus Virus law.invention law Genetics medicine Humans Vector (molecular biology) Molecular Biology Adeno-associated virus Genetic transfer Genetic Therapy Dependovirus beta-Galactosidase Immunohistochemistry Virology Adenoviridae Luminescent Proteins Recombinant DNA Molecular Medicine |
| Zdroj: | Gene Therapy. 6:432-441 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/sj.gt.3300815 |
| Popis: | Gene transfer into the skin is a promising approach to treat inherited or acquired dermatological diseases and systemic monogenic deficiencies. For this purpose, the efficient and sustained gene delivery into keratinocytes is of critical importance. Recombinant adeno-associated virus (rAAV) vectors hold the potential to achieve a long-term gene transfer into various human organs. In order to evaluate this potential for skin gene therapy, human keratinocytes were transduced in vitro with rAAV vectors encoding the reporter genes beta-galactosidase (rAAV/LacZ) or green fluorescent protein (rAAV/GFP). Using rAAV/LacZ at a multiplicity of infection (MOI) of five transducing particles per cell, up to 70% of human keratinocytes were transduced within 48 h. This effect was independent of individual skin donors and different body areas serving as the source for keratinocyte isolation. rAAV had no significant influence on cell viability, but induced a growth arrest in transduced keratinocytes. This growth arrest was overcome by replating cells in fresh media. rAAV/GFP-transduced keratinocytes could be passaged several times, expressed GFP for up to 50 days, and passed the transgene to their daughter cells, suggesting that keratinocyto precursor cells were also transduced. Taken together, the results suggest that rAAV is a promising gene transfer vehicle for skin gene therapy. |
| Databáze: | OpenAIRE |
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