Exploiting the keratin 17 gene promoter to visualize live cells in epithelial appendages of mice.
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.
Bianchi
Nicholas
N
Predoctoral Program in Human Genetics, McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 212105, USA.
Depianto
Daryle
D
McGowan
Kevin
K
Gu
Changhong
C
Coulombe
Pierre A
PA
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D004247:DNA / Q000378:metabolism; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism; D007633:Keratins / Q000235:genetics*; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D012313:RNA / Q000378:metabolism; D012867:Skin / Q000196:embryology*; D013997:Time Factors; D014162:Transfection; D019076:Transgenes
D049452:Green Fluorescent Proteins; D012313:RNA; D007633:Keratins; D004247:DNA; D008156:Luciferases