Advertisement

Transgenesis in Mouse Embryonic Stem Cells

  • Janine van ReeEmail author
  • Wei Zhou
  • Ming Li
  • Jan M. van Deursen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 693)

Abstract

Traditionally, transgenic mice are generated by pronuclear injection of exogenous DNA. This technique has several limitations, including limited control over transgene expression, transgene cytotoxicity, ­promiscuity and silencing, and founder mouse sterility. Here we describe two protocols to generate transgenic mice from ES cell clones carrying stably integrated exogenous DNA with inducible transgene expression.

Key words

Transgenic mice ES cells Inducible expression Cre recombinase Doxycycline 

References

  1. 1.
    Liu, J. and Lobe, C. G. (2007) Cre-conditional expression of constitutively active Notch1 in transgenic mice. Genesis 45, 259–265.PubMedCrossRefGoogle Scholar
  2. 2.
    Novak, A., Guo, C., Yang, W., Nagy, A. and Lobe, C. G. (2000) Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision. Genesis 28, 147–155.PubMedCrossRefGoogle Scholar
  3. 3.
    van Deursen, J. (2003) Gene targeting in mouse embryonic stem cells. Methods Mol. Biol. 209, 145–158.PubMedGoogle Scholar
  4. 4.
    Ramirez-Solis, R., Rivera-Perez, J., Wallace, J. D., Wims, M., Zheng, H. and Bradley, A. (1992) Genomic DNA microextraction: a method to screen numerous samples. Anal. Biochem. 201, 331–335.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Janine van Ree
    • 1
    Email author
  • Wei Zhou
    • 1
  • Ming Li
    • 1
  • Jan M. van Deursen
    • 1
  1. 1.Department of Pediatrics and Adolescent MedicineMayo Clinic College of MedicineRochesterUSA

Personalised recommendations