Skip to main content

Breeding Strategies for Genetically Modified Mice

Part of the Methods in Molecular Biology book series (MIMB,volume 2066)

Abstract

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 is a preview of subscription content, access via your institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • DOI: 10.1007/978-1-4939-9837-1_14
  • Chapter length: 7 pages
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
eBook
USD   109.00
Price excludes VAT (USA)
  • ISBN: 978-1-4939-9837-1
  • Instant PDF download
  • Readable on all devices
  • Own it forever
  • Exclusive offer for individuals only
  • Tax calculation will be finalised during checkout
Softcover Book
USD   149.99
Price excludes VAT (USA)
Hardcover Book
USD   219.99
Price excludes VAT (USA)

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Behringer R (ed) (2014) Manipulating the mouse embryo: a laboratory manual, 4th edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  2. Joyner AL (ed) (2001) Gene targeting: a practical approach, 2nd edn. Oxford University Press, Oxford

    Google Scholar 

  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. https://doi.org/10.1016/j.cell.2013.04.025

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

  4. Singh P, Schimenti JC, Bolcun-Filas E (2015) A mouse geneticist’s practical guide to CRISPR applications. Genetics 199:1–15. https://doi.org/10.1534/genetics.114.169771

    CAS  CrossRef  PubMed  Google Scholar 

  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 27:223–231

    CAS  CrossRef  Google Scholar 

  6. Brinster RL, Chen HY, Warren R, Sarthy A, Palmiter RD (1982) Regulation of metallothionein--thymidine kinase fusion plasmids injected into mouse eggs. Nature 296:39–42

    CAS  CrossRef  Google Scholar 

  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, London

    Google Scholar 

  8. Nagy A (2000) Cre recombinase: the universal reagent for genome tailoring. Genesis 26:99–109

    CAS  CrossRef  Google Scholar 

  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. https://doi.org/10.1038/75973

    CAS  CrossRef  PubMed  Google Scholar 

  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. https://doi.org/10.1016/j.ydbio.2014.06.017

    CAS  CrossRef  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

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

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jan Parker-Thornburg .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and Permissions

Copyright information

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

About this protocol

Verify currency and authenticity via CrossMark

Cite this protocol

Parker-Thornburg, J. (2020). Breeding Strategies for Genetically Modified Mice. In: Larson, M. (eds) Transgenic Mouse. Methods in Molecular Biology, vol 2066. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9837-1_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9837-1_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9836-4

  • Online ISBN: 978-1-4939-9837-1

  • eBook Packages: Springer Protocols