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. 2010 Jun 22;5(6):e11260.
doi: 10.1371/journal.pone.0011260.

Efficient generation of germ line transmitting chimeras from C57BL/6N ES cells by aggregation with outbred host embryos

Marina Gertsenstein  1 , Lauryl M J Nutter, Tammy Reid, Monica Pereira, William L Stanford, Janet Rossant, Andras Nagy
Affiliations

Efficient generation of germ line transmitting chimeras from C57BL/6N ES cells by aggregation with outbred host embryos

Marina Gertsenstein et al. PLoS One. .

Abstract

Genetically modified mouse strains derived from embryonic stem (ES) cells have become essential tools for functional genomics and biomedical research. Large scale mutagenesis projects are producing libraries of mutant C57BL/6 (B6) ES cells to enable the functional annotation of every gene of the mouse genome. To realize the utility of these resources, efficient and accessible methods of generating mutant mice from these ES cells are necessary. Here, we describe a combination of ICR morula aggregation and a chemically-defined culture medium with widely available and accessible components for the high efficiency generation of germline transmitting chimeras from C57BL/6N ES cells. Together these methods will ease the access of the broader biomedical research community to the publicly available B6 ES cell resources.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Representation of fertility and germline transmission status of chimeric mice derived from C57BL/6NTac-C2 targeted clones based on degree of coat colour chimerism.
A. Percentage of mice derived from the same ES cell lines in the same experiments cultured in KOSR+2i or VGB6 prior to aggregation and tested for germline transmission of the ES cell genome. B. Number of mice in each category from A.
See this image and copyright information in PMC

References

    1. Hughes ED, Qu YY, Genik SJ, Lyons RH, Pacheco CD, et al. Genetic variation in C57BL/6 ES cell lines and genetic instability in the Bruce4 C57BL/6 ES cell line. Mamm Genome. 2007;18:549–558. - PubMed
    1. McVicar DW, Winkler-Pickett R, Taylor LS, Makrigiannis A, Bennett M, et al. Aberrant DAP12 signaling in the 129 strain of mice: implications for the analysis of gene-targeted mice. J Immunol. 2002;169:1721–1728. - PubMed
    1. Crawley JN, Belknap JK, Collins A, Crabbe JC, Frankel W, et al. Behavioral phenotypes of inbred mouse strains: implications and recommendations for molecular studies. Psychopharmacology (Berl) 1997;132:107–124. - PubMed
    1. Gerlai R. Gene-targeting studies of mammalian behavior: is it the mutation or the background genotype? Trends Neurosci. 1996;19:177–181. - PubMed
    1. Austin CP, Battey JF, Bradley A, Bucan M, Capecchi M, et al. The knockout mouse project. Nat Genet. 2004;36:921–924. - PMC - PubMed

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