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There has been a significant increase in the use of C57BL/6N-derived ES cells for the production of gene knockout mice. However, the potential for germline transmission (GLT) from chimeras on this genetic background has been observed to be highly variable. Using coat color as an indicator of somatic chimerism to infer the extent of chimeric contribution to the germ cell population, even highly agouti C57BL/6N-derived chimeras can fail to achieve GLT. We investigated the extent to which quantitative PCR genotyping for a marker gene expressed in gene targeted ES cells can be performed on DNA extracted from sperm present in copulatory plugs to determine the contribution of ES cells to the germ cells. We found that an objective assessment of sperm DNA from copulatory plugs combined with a subjective assessment of coat color chimerism can be used to accurately inform the selection of chimeras for breeding that are likely to achieve GLT. These results indicate that, compared to random selection of chimeras, including an analysis of copulatory plugs to set chimeras for breeding can help to reduce costs, minimize time, and facilitate research for projects requiring the production, selection, breeding, and testing of chimeras to generate gene-targeted mice.
Pubmed ID: 23860911 RIS Download
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Center that produces knockout mice and carries out high-throughput phenotyping of each line in order to determine function of every gene in mouse genome. These mice will be preserved in repositories and made available to scientific community representing valuable resource for basic scientific research as well as generating new models for human diseases.
View all literature mentionsNational public repository system for mutant mice. Archives and distributes scientifically valuable spontaneous and induced mutant mouse strains and ES cell lines for use by biomedical research community. Includes breeding/distribution facilities and information coordinating center. Mice strains are cryopreserved, unless live colony must be established. Live mice are supplied from production colony, from colony recovered from cryopreservation, or via micro-injection of cell line into host blastocysts. MMRRC member facilities also develop technologies to improve handling of mutant mice, including advances in assisted reproductive techniques, cryobiology, genetic analysis, phenotyping and infectious disease diagnostics.
View all literature mentionsCenter that imports, archives, maintains, and distributes mutant mouse alleles as live mice, frozen germplasm, stem cells, and molecular vectors for use in biomedical research. The MMRRC Davis receives transgenics, knockouts, and other kinds of mutant mouse lines at no cost to the donor, and after re-derivation and cryopreservation, distributes breeding stock, germplasm, cells, or tissues of genetically-defined and pathogen-free mice for a small fee to requesting investigators.
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