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


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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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.).

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Authors and Affiliations



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).

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