Male and female mice derived from the same embryonic stem cell clone by tetraploid embryo complementation |
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Authors: | Eggan Kevin Rode Anja Jentsch Isabell Samuel Caroline Hennek Thomas Tintrup Hartmut Zevnik Branko Erwin Jennifer Loring Janet Jackson-Grusby Laurie Speicher Michael R Kuehn Ralf Jaenisch Rudolf |
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Institution: | Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, MA 02142, USA. |
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Abstract: | We have devised a general strategy for producing female mice from 39,X0 embryonic stem (ES) cells derived from male cell lines carrying a targeted mutation of interest. We show that the Y chromosome is lost in 2% of subclones from 40,XY ES cell lines, making the identification of targeted 39,X0 subclones a routine procedure. After gene targeting, male and female mice carrying the mutation can be generated by tetraploid embryo complementation from the 40,XY ES cell line and its 39,X0 derivatives. A single intercross then produces homozygous mutant offspring. Because this strategy avoids outcrossing and therefore segregation of mutant alleles introduced into the ES cells, the time and expense required for production of experimental mutant animals from a targeted ES cell clone are substantially reduced. Our data also indicate that ES cells have inherently unstable karyotypes, but this instability does not interfere with production of adult ES cell tetraploid mice. |
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