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Generating Mouse Lines for Lineage Tracing and Knockout Studies

Part of the Methods in Molecular Biology book series (MIMB,volume 1194)

Abstract

In 2007 Capecchi, Evans, and Smithies received the Nobel Prize in recognition for discovering the principles for introducing specific gene modifications in mice via embryonic stem cells, a technology, which has revolutionized the field of biomedical science allowing for the generation of genetically engineered animals. Here we describe detailed protocols based on and developed from these ground-breaking discoveries, allowing for the modification of genes not only to create mutations to study gene function but additionally to modify genes with fluorescent markers, thus permitting the isolation of specific rare wild-type and mutant cell types for further detailed analysis at the biochemical, pathological, and genomic levels.

Key words

  • Mouse
  • Gene targeting
  • Embryonic stem cell culture
  • Pluripotent
  • Chimera
  • Microinjection
  • Germline transmission
  • Cre-recombinase

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Acknowledgements

We are grateful to Siew Lan Lim, Sook Peng Yap, Hsiao Yun Chan, Song Jie, Geraldine Leong, Valerie Tan, Jie Wei Goh, and Manickam Arun Kumar for their helping hands as well as sharing insight and expertise along the way.

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Correspondence to Thomas Lufkin .

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Kraus, P., Sivakamasundari, V., Xing, X., Lufkin, T. (2014). Generating Mouse Lines for Lineage Tracing and Knockout Studies. In: Singh, S., Coppola, V. (eds) Mouse Genetics. Methods in Molecular Biology, vol 1194. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1215-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-1215-5_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1214-8

  • Online ISBN: 978-1-4939-1215-5

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