title: Engineering of Conditional Class I Hdac Knockout Mice and Generation of a Time-Spatial Knockout by a Dual Recombination System.
abstract: The protein sequences of class I HDACs in mice and humans are 96-99 % identical. These highly conserved proteins have crucial roles in biological processes, such as proliferation and development, which is reflected in the lethality that occurs in conventional whole body knockout mice. Therefore, conditional knockouts are inevitable to investigate the functions of class I HDACs in mice. Here, we describe the generation of conditional class I Hdac knockout mice, using Hdac1 as an example. We explain a relatively quick procedure to generate the necessary target vectors by recombination-mediated genetic engineering and gateway techniques. Furthermore, we show how to culture, target, and screen for positively recombined ES cells. Additionally, we present a dual recombination system, which allows the deletion of class I Hdacs at any time by a tamoxifen inducible Cre.
authors.affiliation: Klinikum rechts der Isar, II. Medizinische Klinik, Technische Universität München, Ismaninger Str. 22, 81675, Munich, Germany.
authors.forename: Sieglinde
authors.initials: S
authors.lastname: Bayer
authors.affiliation: Klinikum rechts der Isar, II. Medizinische Klinik, Technische Universität München, Ismaninger Str. 22, 81675, Munich, Germany. matthias.wirth@tum.de.
authors.forename: Matthias
authors.initials: M
authors.lastname: Wirth
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000201:enzymology*; D015139:Blotting, Southern; D003433:Crosses, Genetic; D044127:Epigenesis, Genetic; D005260:Female; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D056284:Histone Deacetylase 1 / Q000172:deficiency / Q000235:genetics*; D059765:Homologous Recombination; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000201:enzymology*; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology
chemical_list: D018841:HSP90 Heat-Shock Proteins; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D013629:Tamoxifen; C045073:Cre recombinase; D019426:Integrases; C484389:Hdac1 protein, mouse; D056284:Histone Deacetylase 1
keywords: Cre/lox; Dual recombination; Flp/FRT; GEMM; HDAC; Knockout; Mouse; Recombination; Tamoxifen; Transgenic