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

Nature Methods volume 6, pages 493–495 (2009)Cite this article

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.

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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.

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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

Author notes

  1. Jennifer L Moran & Kent C Lloyd

    Present address: Present addresses: Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts, USA (J.L.M.) and Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, Massachusetts, USA (K.C.L.).,

Authors and Affiliations

  1. Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK

    Stephen J Pettitt, Qi Liang, Haydn M Prosser, Allan Bradley & William C Skarnes

  2. Center for Comparative Medicine, School of Veterinary Medicine, University of California, Davis, California, USA

    Xin Y Rairdan & Kent C Lloyd

  3. Genetics Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Jennifer L Moran & David R Beier

Authors
  1. Stephen J Pettitt
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  2. Qi Liang
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  4. Jennifer L Moran
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  6. David R Beier
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  7. Kent C Lloyd
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  8. Allan Bradley
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  9. William C Skarnes
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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)

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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

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