Generating Mouse Lines for Lineage Tracing and Knockout Studies

  • Petra Kraus
  • V. Sivakamasundari
  • Xing Xing
  • Thomas LufkinEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1194)


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 



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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Petra Kraus
    • 1
  • V. Sivakamasundari
    • 2
  • Xing Xing
    • 2
  • Thomas Lufkin
    • 1
    Email author
  1. 1.Department of BiologyClarkson UniversityPotsdamUSA
  2. 2.Stem Cell and Developmental Biology, Genome InstituteSingaporeSingapore

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