Transcriptional control in keratinocytes and fibroblasts using synthetic ligands.
The skin is an attractive tissue for regulated target gene expression by virtue of its accessibility to topical regulating stimuli. We have used synthetic ligand-driven intracellular oligomerization to accomplish specific target gene regulation in human skin keratinocytes and fibroblasts. GAL4 DNA binding domains and VP16 transactivation domains, each linked to the FK506 binding protein, were expressed in normal human skin keratinocytes and fibroblasts. These hybrid proteins underwent heterodimerization via the novel intracellular dimerizing agent FK1012 to generate a heterodimeric activator of target gene expression in vitro. Dimeric FK1012, but not monomeric FK506M induced target gene expression in a dose-dependent fashion. FK1012 exerted no detectable nonspecific effects on expression of cutaneous genes and did not alter cellular proliferation kinetics. Controlled oligomerization of hybrid transcription activators offers a potential approach to target gene regulation in cells of normal human skin.
Freiberg
R A
RA
Dermatology Service, VA Palo Alto Health Care System, Palo Alto, California 94304, USA.
Ho
S N
SN
Khavari
P A
PA
D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D015603:Keratinocytes / Q000187:drug effects*; D008024:Ligands; D016559:Tacrolimus / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000187:drug effects*
C084260:FK 1012; D007166:Immunosuppressive Agents; D008024:Ligands; D016559:Tacrolimus