title: Using siRNA knockdown in HaCaT cells to study transcriptional control of epidermal proliferation potential.
abstract: Compared to primary keratinocytes, HaCaT cells are easier to transfect and yet still maintain at least some features of normal epidermal proliferation and differentiation. This chapter describes methods used in our laboratory to maintain HaCaT cells in an undifferentiated state and to use the siRNA technology to efficiently deplete a gene product of interest from these cells. We also provide protocols on how to couple siRNA knockdown with a clonal assay to examine keratinocyte proliferation potential and a luciferase reporter assay to examine promoter regulation.
authors.affiliation: Department of Biological Chemistry, School of Medicine, University of California, Irvine, Irvine, CA, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Wells
authors.forename: Xing
authors.initials: X
authors.lastname: Dai
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D015603:Keratinocytes / Q000166:cytology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering; D014158:Transcription, Genetic
chemical_list: D034741:RNA, Small Interfering; D008156:Luciferases