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Generation of Genetically Modified Organotypic Skin Cultures Using Devitalized Human Dermis
Published on: December 14, 2015
Defining the regulatory factors required for epidermal gene expression
S Sinha1, L Degenstein, C Copenhaver
1Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois 60637, USA.
Molecular and Cellular Biology
|March 14, 2000
Summary
Keratin 14 (K14) gene regulation in epidermis was studied using transgenic mice and cell cultures. Key regulatory elements were identified within the K14 gene
Area of Science:
- Molecular Biology
- Genetics
- Dermatology
Background:
- Keratins K5 and K14 are crucial for stratified epithelia.
- Their high-level, tissue-specific transcription offers insight into epidermis regulation.
Purpose of the Study:
- To identify regulatory mechanisms of epidermis-specific transcription for the K14 gene.
- To elucidate the role of regulatory elements in keratinocyte gene expression.
Main Methods:
- Analysis of DNase I-hypersensitive sites (HSs) in K14 5' regulatory sequences.
- In vivo transgenic approaches and in vitro keratinocyte culture.
- Electrophoretic mobility shift and mutational assays.
Main Results:
- Identified a 700-bp regulatory domain containing HSsII and -III critical for K14 expression.
- A 125-bp fragment with HSsII demonstrated enhancer activity.
- AP-1, ets, and AP-2 transcription factors are involved in K14 regulation within HSsII.
Conclusions:
- A novel enhancer region regulates K14 transcription in keratinocytes.
- Specific transcription factors (ets, AP-2) within HSsII are key for epidermal specificity.
- Further investigation is needed to fully restrict activity to keratinocytes in vivo.
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