title: Generation of transgenic mice on an NOD/SCID background using the conventional microinjection technique.
abstract: Humanized mice, which refers to immunodeficient mice repopulated with the human immune system, are powerful tools for study in the field of immunology. It has been difficult, however, to generate these transgenic (Tg) mice directly from such strains as the NOD/SCID mouse. In this study, we describe a method developed by us for the generation of Tg mice on an NOD/SCID background. First, we obtained fertilized eggs efficiently by means of in vitro fertilization (IVF); then, we attempted to generate CAG-EGFP Tg mice on an NOD/SCID background, finding that delayed timing of the microinjection after the IVF improved the time to development of the two-cell-stage embryos and the obtainment of newborns. We successfully generated Tg mice and confirmed the germ-line transmission in the offspring. In conclusion, we established a novel system for directly generating transgenic mice on an NOD/SCID background. This novel system is expected to allow improved efficiency of the generation of humanized mice.
authors.affiliation: Department of Metabolic Diseases, The University of Tokyo, Tokyo, Japan.
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Kumagai
authors.forename: Naoto
authors.initials: N
authors.lastname: Kubota
authors.forename: Toshiki I
authors.initials: TI
authors.lastname: Saito
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Sasako
authors.forename: Rumi
authors.initials: R
authors.lastname: Takizawa
authors.forename: Katsuko
authors.initials: K
authors.lastname: Sudo
authors.forename: Mineo
authors.initials: M
authors.lastname: Kurokawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Kadowaki
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D016688:Mice, Inbred NOD / Q000235:genetics* / Q000276:immunology*; D016513:Mice, SCID / Q000235:genetics* / Q000276:immunology*; D008822:Mice, Transgenic / Q000235:genetics* / Q000276:immunology*; D008845:Microinjections / Q000379:methods; D011247:Pregnancy
chemical_list: D017931:DNA Primers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins