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Molecular and Cellular Biology, August 2005, p. 7249-7259, Vol. 25, No. 16
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.16.7249-7259.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Exploiting the Keratin 17 Gene Promoter To Visualize Live Cells in Epithelial Appendages of Mice

Nicholas Bianchi,¹ Daryle DePianto,² Kevin McGowan,^2, Changhong Gu,² and Pierre A. Coulombe^2,3*

Predoctoral Program in Human Genetics, McKusick-Nathans Institute of Genetic Medicine,¹ Department of Biological Chemistry,² Department of Dermatology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205³

Received 10 February 2005/ Returned for modification 17 March 2005/ Accepted 19 May 2005

Keratin genes afford, given their large number (>50) and differential regulation, a unique opportunity to study the mechanisms underlying specification and differentiation in epithelia of higher metazoans. Moreover, the small size and regulation in cis of many keratin genes enable the use of their regulatory sequence to achieve targeted gene expression in mice. Here we show that 2 kilobases of 5' upstream region from the mouse keratin 17 gene (mK17) confers expression of green fluorescent protein (GFP) in major epithelial appendages of transgenic mice. Like that of mK17, onset of [mK17 5']-GFP reporter expression coincides with the appearance of ectoderm-derived epithelial appendages during embryonic development. In adult mice, [mK17 5']-GFP is appropriately regulated within hair, nail, glands, and oral papilla. Tracking of GFP fluorescence allows for the visualization of growth cycle-related changes in hair follicles, and the defects engendered by the hairless mutation, in live skin tissue. Deletion of an internal 48-bp interval, which encompasses a Gli-responsive element, from this promoter results in loss of GFP fluorescence in most appendages in vivo, suggesting that sonic hedgehog participates in K17 regulation. The compact mK17 gene promoter provides a novel tool for appendage-preferred gene expression and manipulation in transgenic mice.

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* Corresponding author. Mailing address: Department of Biological Chemistry, Johns Hopkins University School of Medicine, 725 N. Wolfe Street, Baltimore, MD 212105. Phone: (410) 614-0510. Fax: (410) 614-7567. E-mail: coulombe{at}jhmi.edu.

Supplemental material for this article may be found at http://mcb.asm.org/.

Present address: ExonHit Therapeutics, 217 Perry Parkway, Gaithersburg, MD 20877.

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Molecular and Cellular Biology, August 2005, p. 7249-7259, Vol. 25, No. 16
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.16.7249-7259.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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