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Engineered human skin model using poly(ADP-ribose) polymerase antisense expression shows a reduced response to DNA
D S Rosenthal1, T B Shima, G Celli
1Department of Biochemistry and Molecular Biology, Georgetown University School of Medicine, Washington, D.C. 20007, USA.
The Journal of Investigative Dermatology
|July 1, 1995
Summary
Researchers developed a novel model to study Poly(ADP-ribose) polymerase (PADPRP) in skin cells. This system allows selective reduction of PADPRP, aiding research into DNA repair mechanisms in the epidermis.
Area of Science:
- Biochemistry
- Dermatology
- Molecular Biology
Background:
- Poly(ADP-ribose) polymerase (PADPRP) is crucial for cellular responses to DNA damage.
- The skin epidermis is frequently exposed to DNA-damaging agents.
- Understanding PADPRP's role in epidermal maintenance is vital.
Purpose of the Study:
- To develop a model system for studying Poly(ADP-ribose) polymerase (PADPRP) in human keratinocytes.
- To investigate the function of PADPRP in epidermal maintenance and DNA repair.
Main Methods:
- Stable transfection of human keratinocytes with antisense RNA targeting PADPRP under an inducible promoter.
- Induction of antisense RNA to selectively reduce PADPRP mRNA, protein, and activity.
- Grafting of transfected keratinocytes onto nude mice to form reconstituted human skin for in vivo studies.
Main Results:
- Antisense RNA induction effectively lowered PADPRP levels and activity in cultured keratinocytes.
- Reduced PADPRP impaired the cells' ability to synthesize poly(ADP-ribose) in response to DNA damage.
- Grafted skin formed histologically normal tissue, and PADPRP levels were inducible in vivo.
Conclusions:
- A functional model system was established to manipulate PADPRP levels in human keratinocytes and reconstituted epidermis.
- This model is a valuable tool for investigating the role of PADPRP and DNA repair in epidermal biology.