title: Rapid selection of XO embryonic stem cells using Y chromosome-linked GFP transgenic mice.
abstract: We developed a transgenic mouse line with Y chromosome-linked green fluorescent protein expressing transgenes (Y-GFP) by the conventional microinjection into the pronucleus of C57BL/6J fertilized oocytes. Embryonic stem (ES) cells derived from Y-GFP mice enabled not only sexing but also the identification of 39, XO karyotype by the lack of Y chromosome. Actually, when fluorescence activated cell sorting (FACS) was applied to Y-GFP ES cells, non-fluorescent ES cells were conveniently collected and showed the lack of Y chromosome by PCR genotyping and Southern blot analysis. FACS analysis revealed Y chromosome loss occurred at 2.9 % of 40, XY ES cells after five passages. These Y-GFP ES cells are potentially applicable to reduce the time, cost and effort needed to generate the gene-targeted mice by the production of male and female mice derived from the same ES cell clone.
authors.affiliation: Department of Biochemistry, Jichi Medical University, 3311-1, Yakushiji, Shimotsuke, Tochigi, 329-0498, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Yasumitsu
authors.initials: Y
authors.lastname: Nagao
authors.forename: Kenji
authors.initials: K
authors.lastname: Kuroiwa
authors.forename: Yoji
authors.initials: Y
authors.lastname: Hakamata
authors.forename: Masaru
authors.initials: M
authors.lastname: Ichida
authors.forename: Fumiko
authors.initials: F
authors.lastname: Saito-Ohara
authors.forename: Kaoru
authors.initials: K
authors.lastname: Tominaga
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Endo
mesh_terms: D059786:Abnormal Karyotype; D000818:Animals; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D018014:Gene Transfer Techniques; D050173:Genes, Y-Linked / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction; D019076:Transgenes / Q000235:genetics*
chemical_list: D017931:DNA Primers; D049452:Green Fluorescent Proteins