title: Improved experimental procedures for achieving efficient germ line transmission of nonobese diabetic (NOD)-derived embryonic stem cells.
abstract: The manipulation of a specific gene in NOD mice, the best animal model for insulin-dependent diabetes mellitus (IDDM), must allow for the precise characterization of the functional involvement of its encoded molecule in the pathogenesis of the disease. Although this has been attempted by the cross-breeding of NOD mice with many gene knockout mice originally created on the 129 or C57BL/6 strain background, the interpretation of the resulting phenotype(s) has often been confusing due to the possibility of a known or unknown disease susceptibility locus (e.g., Idd locus) cosegregating with the targeted gene from the diabetes-resistant strain. Therefore, it is important to generate mutant mice on a pure NOD background by using NOD-derived embryonic stem (ES) cells. By using the NOD ES cell line established by Nagafuchi and colleagues in 1999 (FEBS Lett., 455, 101-104), the authors reexamined various conditions in the context of cell culture, DNA transfection, and blastocyst injection, and achieved a markedly improved transmission efficiency of these NOD ES cells into the mouse germ line. These modifications will enable gene targeting on a "pure" NOD background with high efficiency, and contribute to clarifying the physiological roles of a variety of genes in the disease course of IDDM.
authors.affiliation: Center for Immunology, University of Texas Southwestern Medical Center, Dallas, TX 75390-9093, USA.
authors.forename: Satoko
authors.initials: S
authors.lastname: Arai
authors.forename: Christina
authors.initials: C
authors.lastname: Minjares
authors.forename: Seiho
authors.initials: S
authors.lastname: Nagafuchi
authors.forename: Toru
authors.initials: T
authors.lastname: Miyazaki
mesh_terms: D000818:Animals; D001755:Blastocyst; D002417:Cattle / Q000097:blood; D002460:Cell Line; D004247:DNA; D004622:Embryo, Mammalian / Q000166:cytology; D005312:Fetal Blood; D018014:Gene Transfer Techniques; D005854:Germ Cells; D007267:Injections; D051379:Mice; D016688:Mice, Inbred NOD / Q000196:embryology*; D033581:Stem Cell Transplantation; D013234:Stem Cells; D014162:Transfection; D018183:Transplantation Chimera
chemical_list: D004247:DNA