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Male and female mice derived from the same embryonic stem cell clone by tetraploid embryo complementation


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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Figure 1: Standard and accelerated production of mutant mice from ES cells.
Figure 2: Isolation of targeted 39,X0 subclones from 40,XY ES cell lines.
Figure 3: Mutant female mice produced from 39,X0 ES cells by tetraploid embryo complementation when crossed to ES cell–tetraploid males produce homozygous mutant offspring.

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We would like to thank D. Page, W.M. Rideout, C. Beard, K. Hochedlinger, A. Bortvin, M. Rios, and D. Menke for helpful discussions. This work was supported in part by NIH grants 5-R35-CA44339 and RO1-CA84198 to R. Jaenisch. I. Jentsch received a stipend from TILL I.D., Gräfeling, Germany.

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Correspondence to Rudolf Jaenisch.

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A.R., C.S., T.H., H.T., B.Z., and R.K. are employed by Artemis Pharmaceuticals, Cologne, and will use some of the results reported here for their ongoing commercial interests. The remaining authors are all at academic institutions that are supported by public funds and have no competing interests in this research.

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Eggan, K., Rode, A., Jentsch, I. et al. Male and female mice derived from the same embryonic stem cell clone by tetraploid embryo complementation. Nat Biotechnol 20, 455–459 (2002).

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