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

  • ️Jaenisch, Rudolf
  • ️Wed May 01 2002

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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References

  1. Gu, H., Marth, J.D., Orban, P.C., Mossmann, H. & Rajewsky, K. Deletion of a DNA polymerase β gene segment in T cells using cell type-specific gene targeting. Science 265, 103–106 (1994).

    Article  CAS  Google Scholar 

  2. Rodriguez, C.I. et al. High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. Nat. Genet. 25, 139–140 (2000).

    Article  CAS  Google Scholar 

  3. Lakso, M. et al. Targeted oncogene activation by site-specific recombination in transgenic mice. Proc. Natl. Acad. Sci. USA 89, 6232–6236 (1992).

    Article  CAS  Google Scholar 

  4. Minichiello, L. et al. Essential role for TrkB receptors in hippocampus-mediated learning. Neuron 24, 401–414 (1999).

    Article  CAS  Google Scholar 

  5. Shibata, H. et al. Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene. Science 278, 120–123 (1997).

    Article  CAS  Google Scholar 

  6. Eggan, K. et al. Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraploid embryo complementation. Proc. Natl. Acad. Sci. USA 98, 6209–6214 (2001).

    Article  CAS  Google Scholar 

  7. Nagy, A., Rossant, J., Nagy, R., Abramow-Newerly, W. & Roder, J.C. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc. Natl. Acad. Sci. USA 90, 8424–8428 (1993).

    Article  CAS  Google Scholar 

  8. Cattanach, B. X0 Mice. Genet. Res. 3, 487–490 (1962).

    Article  Google Scholar 

  9. Lamar, E.E. & Palmer, E. Y-encoded, species-specific DNA in mice: evidence that the Y chromosome exists in two polymorphic forms in inbred strains. Cell 37, 171–177 (1984).

    Article  CAS  Google Scholar 

  10. Passemato, R., Eggan, K., Moeller, B., Jaenisch, R. & Jackson-Grusby, L. Flp recombinase regulated lacz expression at the ROSA26 locus. Genesis. 32, 184–186 (2002).

    Article  Google Scholar 

  11. Jentsch, I., Adler, I.D., Carter, N.P. & Speicher, M.R. Karyotyping mouse chromosomes by multiplex-FISH (M-FISH). Chromosome Res. 9, 211–214 (2001).

    Article  CAS  Google Scholar 

  12. Brook, J.D. X-chromosome segregation, maternal age and aneuploidy in the X0 mouse. Genet. Res. 41, 85–95 (1983).

    Article  CAS  Google Scholar 

  13. Al-Shawi, R. et al. The herpes simplex virus type 1 thymidine kinase is expressed in the testes of transgenic mice under the control of a cryptic promoter. Mol. Cell. Biol. 11, 4207–4216 (1991).

    Article  CAS  Google Scholar 

  14. Wang, P.J., McCarrey, J.R., Yang, F. & Page, D.C. An abundance of X-linked genes expressed in spermatogonia. Nat. Genet. 27, 422–426 (2001).

    Article  Google Scholar 

  15. Marahrens, Y., Panning, B., Dausman, J., Strauss, W. & Jaenisch, R. Xist-deficient mice are defective in dosage compensation but not spermatogenesis. Genes. Dev. 11, 156–166 (1997).

    Article  CAS  Google Scholar 

  16. Liu, X. et al. Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev. Dyn. 209, 85–91 (1997).

    Article  CAS  Google Scholar 

  17. Hogan, B. et al. Isolation, culture, and manipulation of embryonic stem cells. In Manipulating the Mouse Embryo. (eds Hogan, B., Beddington, R., Constantini, F. and Lacy, E.) 253–289 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 1994).

    Google Scholar 

  18. Bishop, C.E. & Hatat, D. Molecular cloning and sequence analysis of a mouse Y chromosome RNA transcript expressed in the testis. Nucleic Acids Res. 15, 2959–2969 (1987).

    Article  CAS  Google Scholar 

  19. Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat. Genet. 21, 70–71 (1999).

    Article  CAS  Google Scholar 

  20. Laird, P.W. et al. Simplified mammalian DNA isolation procedure. Nucleic Acids Res. 19, 4293 (1991).

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Acknowledgements

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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Authors and Affiliations

  1. Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, 02142, MA, USA

    Kevin Eggan, Jennifer Erwin, Janet Loring, Laurie Jackson-Grusby & Rudolf Jaenisch

  2. Department of Biology, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, 02142, MA, USA

    Kevin Eggan, Jennifer Erwin & Rudolf Jaenisch

  3. Artemis Pharmaceuticals GmbH, Neurather Ring 1, Cologne, D-51063, Germany

    Anja Rode, Caroline Samuel, Thomas Hennek, Hartmut Tintrup, Branko Zevnik & Ralf Kuehn

  4. Institute of Human Genetics, Technical University, Munich, Germany

    Isabell Jentsch & Michael R. Speicher

  5. Institute of Human Genetics, GSF National Research Center, Munich, Germany

    Isabell Jentsch & Michael R. Speicher

Authors

  1. Kevin Eggan

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  2. Anja Rode

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  3. Isabell Jentsch

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  4. Caroline Samuel

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  5. Thomas Hennek

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  6. Hartmut Tintrup

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  7. Branko Zevnik

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  8. Jennifer Erwin

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  9. Janet Loring

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  10. Laurie Jackson-Grusby

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  11. Michael R. Speicher

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  12. Ralf Kuehn

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  13. Rudolf Jaenisch

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Corresponding author

Correspondence to Rudolf Jaenisch.

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Competing interests

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). https://doi.org/10.1038/nbt0502-455

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  • Received: 06 December 2001

  • Accepted: 13 February 2002

  • Issue Date: 01 May 2002

  • DOI: https://doi.org/10.1038/nbt0502-455

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