Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Agouti C57BL/6N embryonic stem cells for mouse genetic resources

Abstract

We report the characterization of a highly germline competent C57BL/6N mouse embryonic stem cell line, JM8. To simplify breeding schemes, the dominant agouti coat color gene was restored in JM8 cells by targeted repair of the C57BL/6 nonagouti mutation. These cells provide a robust foundation for large-scale mouse knockout programs that aim to provide a public resource of targeted mutations in the C57BL/6 genetic background.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Properties and origin of Agouti JM8 C57BL/6N embryonic stem cells.
Figure 2: Coat color of chimeric mice and their offspring from injections of JM8 and JM8A3 cells into blastocysts from several common mouse strains.

References

  1. Mouse Genome Sequencing Consortium. Nature 420, 520–562 (2002).

  2. International Mouse Knockout Consortium. Cell 128, 9–13 (2007).

  3. Kontgen, F., Suss, G., Stewart, C., Steinmetz, M. & Bluethmann, H. Int. Immunol. 5, 957–964 (1993).

    Article  CAS  Google Scholar 

  4. Auerbach, W. et al. Biotechniques 29, 1024–1032 (2000).

    Article  CAS  Google Scholar 

  5. Schuster-Gossler, K. et al. Biotechniques 31, 1022–1026 (2001).

    Article  CAS  Google Scholar 

  6. Cheng, J., Dutra, A., Takesono, A., Garrett-Beal, L. & Schwartzberg, P. Genesis 39, 100–104 (2004).

    Article  Google Scholar 

  7. Shimizukawa, R. et al. Genesis 42, 47–52 (2005).

    Article  CAS  Google Scholar 

  8. Hughes, E. et al. Mamm. Genome 18, 549–558 (2007).

    Article  CAS  Google Scholar 

  9. Keskintepe, L., Norris, K., Pacholczyk, G., Dederscheck, S. & Eroglu, A. Transgenic Res. 16, 751–758 (2007).

    Article  CAS  Google Scholar 

  10. Hansen, G.M. et al. Genome Res. 18, 1670–1679 (2008).

    Article  CAS  Google Scholar 

  11. Nagy, A., Gertesenstein, M., Vintersten, K. & Behringer, R. Manipulating the Mouse Embryo: A Laboratory Manual 3rd ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2003).

    Google Scholar 

  12. Stiedl, O. et al. Behav. Brain Res. 104, 1–12 (1999).

    Article  CAS  Google Scholar 

  13. Khisti, R., Wolstenholme, J., Shelton, K. & Miles, M. Alcohol 40, 119–126 (2006).

    Article  CAS  Google Scholar 

  14. Liang, Q., Conte, N., Skarnes, W. & Bradley, A. Proc. Natl. Acad. Sci. USA 105, 17453–17456 (2008).

    Article  CAS  Google Scholar 

  15. Bultman, S. et al. Genes Dev. 8, 481–490 (1994).

    Article  CAS  Google Scholar 

  16. McMahon, A. & Bradley, A. Cell 62, 1073–1085 (1990).

    Article  CAS  Google Scholar 

  17. Liu, P., Zhang, H., McLellan, A., Vogel, H. & Bradley, A. Genetics 150, 1155–1168 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Court, D.L. et al. Gene 315, 63–69 (2003).

    Article  CAS  Google Scholar 

  19. Farley, F., Soriano, P., Steffen, L. & Dymecki, S. Genesis 28, 106–110 (2000).

    Article  CAS  Google Scholar 

  20. Dickie, M. J. Hered. 60, 20–25 (1969).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Qin, F. Law, L. Delaney, J. Meneses, T. Creek, H. Tharagonnet and A. Beasley for help with embryonic stem cell culture; N. Adams, J. White, R. Ramirez-Solis and the Wellcome Trust Sanger Institute microinjection team for blastocyst injections and mouse breeding; and N. Conte and M. Storer for help with CGH. This work was funded by the Wellcome Trust Sanger Institute (WT077187), grants from the National Institutes of Health (KOMP, U01-HG004080 to W.C.S., K.C.L. and A.B.; U01-42430 to D.R.B.) and a grant from the Sixth Framework Programme of the EU (EUCOMM, to W.C.S. and A.B.).

Author information

Authors and Affiliations

Authors

Contributions

S.J.P. repaired the Agouti allele, analyzed the JM8A3 subline by CGH, microinjection and test crosses, and wrote the paper; H.M.P. and A.B. designed agouti targeting strategy and supervised experiments; Q.L. analyzed JM8 subclones by CGH; X.Y.R. and K.C.L. derived the parental JM8 line; J.L.M. and D.R.B. discovered and genotyped SNPs; A.B. designed experiments and assisted in writing the paper; W.C.S. derived JM8.F and JM8.N subclones, designed and supervised targeting and microinjection experiments and wrote the paper.

Corresponding author

Correspondence to William C Skarnes.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–2, Supplementary Table 1, Supplementary Note (PDF 1204 kb)

Supplementary Data 1

Microinjection and breeding data for JM8 subclones (XLS 18 kb)

Supplementary Data 2

Microinjection and breeding data for targeted subclones (XLS 111 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Pettitt, S., Liang, Q., Rairdan, X. et al. Agouti C57BL/6N embryonic stem cells for mouse genetic resources. Nat Methods 6, 493–495 (2009). https://doi.org/10.1038/nmeth.1342

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmeth.1342

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing