High-efficiency gene transfer and pharmacologic selection of genetically engineered human keratinocytes.
Low efficiencies of gene transfer to somatic cells have frustrated therapeutic gene delivery efforts in a wide array of tissues including the skin. Production of populations of keratinocytes in which all cells contain the desired therapeutic gene may be important in future genetic therapies. This may be the case in disorders such as epidermolysis bullosa and ichthyosis, where a failure to correct the vast majority of cells within tissue could perpetuate central disease features such as skin fragility and defective barrier function. We have refined retroviral gene transfer parameters to achieve significant improvements in gene delivery efficiencies to human keratinocytes compared to those previously reported. We have also generated retroviral vectors that allow rapid pharmacologic selection of human keratinocytes without interfering with the potential of these cells to regenerate epidermis in vivo--we determined that blasticidin is superior to the commonly used neomycin. The combined capabilities for efficient retroviral gene transfer and effective pharmacologic selection allow production of entirely engineered populations of human keratinocytes for use in future efforts to achieve effective cutaneous gene delivery.[1]References
- High-efficiency gene transfer and pharmacologic selection of genetically engineered human keratinocytes. Deng, H., Choate, K.A., Lin, Q., Khavari, P.A. BioTechniques (1998) [Pubmed]
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