Breeding Strategies for Genetically Modified Mice

  • Jan Parker-ThornburgEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2066)


Genetically modified mice can be generated using a variety of methods. Depending on the method used, the breeding strategy must be modified not only for the initial founding generation of mouse, but to establish individual mouse lines as well. Transgenic founder mice are each unique and must be outcrossed to initially establish the line. Mice that have been targeted using embryonic stem cells will need to be tested for germ line transmission of the modified gene prior to establishing a line. Mice that have been targeted using CRISPR/Cas9 gene-editing endonucleases will often be mosaic, and thus, also require testing for germ line transmission of the mutation of interest.

Key words

Transgenic Knock-out mice Knock-in mice CRISPR CRISPR/Cas9 Germ line transmission Chimera Mosaic 



This work was supported by NCI grant #CA016672 (GEMF), a Cancer Center Support Grant to MD Anderson Cancer Center and NCI grant #R50CA211121.


  1. 1.
    Behringer R (ed) (2014) Manipulating the mouse embryo: a laboratory manual, 4th edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  2. 2.
    Joyner AL (ed) (2001) Gene targeting: a practical approach, 2nd edn. Oxford University Press, OxfordGoogle Scholar
  3. 3.
    Wang H, Yang H, Chivalila CS, Dawlaty MM, Cheng AW, Zheng F, Jaenisch R (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153:910–918. Scholar
  4. 4.
    Singh P, Schimenti JC, Bolcun-Filas E (2015) A mouse geneticist’s practical guide to CRISPR applications. Genetics 199:1–15. Scholar
  5. 5.
    Brinster RL, Chen HY, Trumbauer M, Senear AW, Warren R, Palmiter RD (1981) Somatic expression of herpes thymidine kinase in mice following injection of a fusion gene into eggs. Cell 27:223–231CrossRefGoogle Scholar
  6. 6.
    Brinster RL, Chen HY, Warren R, Sarthy A, Palmiter RD (1982) Regulation of metallothionein--thymidine kinase fusion plasmids injected into mouse eggs. Nature 296:39–42CrossRefGoogle Scholar
  7. 7.
    Williams E, Auerbach W, DeChiara TM, Gertsenstein M (2011) Combining ES cells with embryos. In: Pease S, Saunders TL (eds) Advanced protocols for animal transgenesis. Springer, LondonGoogle Scholar
  8. 8.
    Nagy A (2000) Cre recombinase: the universal reagent for genome tailoring. Genesis 26:99–109CrossRefGoogle Scholar
  9. 9.
    Rodriguez CI, Buchholz F, Galloway J, Sequerra R, Kasper J, Ayala R, Stewart AF, Dymecki SM (2000) High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP. Nat Genet 25:139–140. Scholar
  10. 10.
    Yen ST, Zhang M, Deng JM, Usman SJ, Smith CN, Parker-Thornburg J, Swinton PG, Martin JF, Behringer RR (2014) Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes. Dev Biol 393:3–9. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Genetically Engineered Mouse Facility, Department of GeneticsMD Anderson Cancer CenterHoustonUSA

Personalised recommendations