Rapid Selection Of XO Embryonic Stem Cells Using Y Chromosome ...

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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.

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References

1. 1. Nat Genet. 1998 Jul;19(3):220-2 - PubMed
2. 1. Gene. 1991 Dec 15;108(2):193-9 - PubMed
3. 1. Nat Methods. 2011 Oct 23;8(12):1071-7 - PubMed
4. 1. Nature. 2008 May 22;453(7194):519-23 - PubMed
5. 1. Nucleic Acids Res. 1987 Apr 10;15(7):2959-69 - PubMed

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