title: Rho GTPase knockout induction in primary keratinocytes from adult mice.
abstract: Primary keratinocytes are an important tool to investigate the molecular mechanism underlying the skin phenotype of mice with null mutations in Rho GTPase genes. If the RhoA gene deletion is conditional, the knockout can be induced in vitro by transfection with cre-IRES-GFP and sorting for GFP positive cells by flow cytometry. Such in vitro knockout will allow determining the cell autonomous functions of the Rho GTPase, independent of any in vivo interactions. Using the same method, also other expression vectors or knockdown constructs can be introduced into primary mouse keratinocytes.
authors.affiliation: Molecular Pathology Section, Department of Biomedical Sciences, BRIC, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Esben
authors.initials: E
authors.lastname: Pedersen
authors.forename: Astrid
authors.initials: A
authors.lastname: Basse
authors.forename: Tine
authors.initials: T
authors.lastname: Lefever
authors.forename: Karine
authors.initials: K
authors.lastname: Peyrollier
authors.forename: Cord
authors.initials: C
authors.lastname: Brakebusch
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods; D005434:Flow Cytometry; D055786:Gene Knockout Techniques; D015603:Keratinocytes / Q000201:enzymology*; D051379:Mice; D061251:Primary Cell Culture / Q000379:methods; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000235:genetics*
chemical_list: D020741:rho GTP-Binding Proteins