Abstract
Genetically modified mice by means of homologous recombination in embryonic stem (ES) cells are generated by injection of manipulated ES cells into recipient blastocysts. The injected blastocysts, following reintroduction into recipient foster mice, will produce chimeric mice in which the manipulated ES cells populate the germline and transmit the induced mutation to the offspring. Crossing of the chimeras’ offspring bearing the targeted mutation in heterozygosis will ultimately produce mice homozygous for the specific genetic mutation. Here we describe the steps and procedures required to generate the chimeric mice leading to the transfer of a genetic mutation to the mouse germline.
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Acknowledgments
We thank Eileen Southon for critical reading of the manuscript. This research was supported by the Intramural Research Program of the NIH, National Cancer Institute. The contents of this publication do not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. The Center for Cancer Research, NCI-Frederick has filed an Animal Welfare Assurance with the Office for Protection from Research Risks (OPRR). The protocols herein described have been approved by the NCI-Frederick Institutional Animal Care and Use Committee.
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Reid, S.W., Tessarollo, L. (2009). Isolation, Microinjection and Transfer of Mouse Blastocysts. In: Wurst, W., Kühn, R. (eds) Gene Knockout Protocols. Methods in Molecular Biology, vol 530. Humana Press. https://doi.org/10.1007/978-1-59745-471-1_14
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DOI: https://doi.org/10.1007/978-1-59745-471-1_14
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