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TALENs pp 229-243 | Cite as

Genome Editing in Mice Using TALE Nucleases

  • Benedikt Wefers
  • Christina Brandl
  • Oskar Ortiz
  • Wolfgang WurstEmail author
  • Ralf KühnEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1338)

Abstract

Gene engineering for generating targeted mouse mutants is a key technology for biomedical research. Using TALENs as sequence-specific nucleases to induce targeted double-strand breaks, the mouse genome can be directly modified in zygotes in a single step without the need for embryonic stem cells. By embryo microinjection of TALEN mRNAs and targeting vectors, knockout and knock-in alleles can be generated fast and efficiently. In this chapter we provide protocols for the application of TALENs in mouse zygotes.

Key words

Knock-in Knockout One-cell embryo Gene targeting TALEN Mouse 

Notes

Acknowledgements

This work was supported by the European Union within the EUCOMM project (LSHG-CT-2005-018931, to W.W.), by the German Ministry of Education and Research within the TAL-CUT (03 V0261) and DIGTOP project (01GS0858) (to W.W. and R.K.) of the VIP and NGFN-Plus program.

References

  1. 1.
    Capecchi MR (2005) Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century. Nat Rev Genet 6:507–12CrossRefPubMedGoogle Scholar
  2. 2.
    Meyer M, de Angelis MH, Wurst W et al (2010) Gene targeting by homologous recombination in mouse zygotes mediated by zinc-finger nucleases. Proc Natl Acad Sci U S A 107:15022–6PubMedCentralCrossRefPubMedGoogle Scholar
  3. 3.
    Carbery ID, Ji D, Harrington A et al (2010) Targeted genome modification in mice using zinc-finger nucleases. Genetics 186:451–9PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Boch J, Scholze H, Schornack S et al (2009) Breaking the code of DNA binding specificity of TAL-type III effectors. Science 326:1509–1512CrossRefPubMedGoogle Scholar
  5. 5.
    Moscou MJ, Bogdanove AJ (2009) A simple cipher governs DNA recognition by TAL effectors. Science 326:1501CrossRefPubMedGoogle Scholar
  6. 6.
    Christian M, Cermak T, Doyle EL et al (2010) Targeting DNA double-strand breaks with TAL effector nucleases. Genetics 186:757–761PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Miller JC, Tan S, Qiao G et al (2011) A TALE nuclease architecture for efficient genome editing. Nat Biotechnol 29:143–148CrossRefPubMedGoogle Scholar
  8. 8.
    Heyer W-D, Ehmsen KT, Liu J (2010) Regulation of homologous recombination in eukaryotes. Annu Rev Genet 44:113–139PubMedCentralCrossRefPubMedGoogle Scholar
  9. 9.
    Lieber MR (2010) The mechanism of double-strand DNA break repair by the nonhomologous DNA end-joining pathway. Annu Rev Biochem 79:181–211PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Sung YH, Baek I-J, Kim DH et al (2013) Knockout mice created by TALEN-mediated gene targeting. Nat Biotechnol 31:23–24CrossRefPubMedGoogle Scholar
  11. 11.
    Kato T, Miyata K, Sonobe M et al (2013) Production of Sry knockout mouse using TALEN via oocyte injection. Sci Rep 3:3136PubMedCentralPubMedGoogle Scholar
  12. 12.
    Matsubara Y, Kato T, Kashimada K et al (2015) TALEN-mediated gene disruption on Y Chromosome reveals Critical Role of EIF2S3Y in Mouse Spermatogenesis. Stem Cells Dev 24(10):1164–70CrossRefPubMedGoogle Scholar
  13. 13.
    Kasparek P, Krausova M, Haneckova R et al (2014) Efficient gene targeting of the Rosa26 locus in mouse zygotes using TALE nucleases. FEBS Lett 588:3982–3988CrossRefPubMedGoogle Scholar
  14. 14.
    Wefers B, Meyer M, Ortiz O et al (2013) Direct production of mouse disease models by embryo microinjection of TALENs and oligodeoxynucleotides. Proc Natl Acad Sci U S A 110:3782–3787PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Panda SK, Wefers B, Ortiz O et al (2013) Highly efficient targeted mutagenesis in mice using TALENs. Genetics 195:703–713PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Brandl C, Ortiz O, Röttig B et al (2014) Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one-cell mouse embryos. FEBS Open Bio 5:26–35PubMedCentralCrossRefPubMedGoogle Scholar
  17. 17.
    Davies B, Davies G, Preece C et al (2013) Site specific mutation of the Zic2 locus by microinjection of TALEN mRNA in mouse CD1, C3H and C57BL/6J oocytes. PLoS One 8, e60216PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Qiu Z, Liu M, Chen Z et al (2013) High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases. Nucleic Acids Res 41, e120PubMedCentralCrossRefPubMedGoogle Scholar
  19. 19.
    Ma K, Wang J, Shen B et al (2013) Efficient targeting of FATS at a common fragile site in mice through TALEN-mediated double-hit genome modification. Biotechnol Lett 36(3):471–9CrossRefPubMedGoogle Scholar
  20. 20.
    Takada S, Sato T, Ito Y et al (2013) Targeted Gene Deletion of miRNAs in Mice by TALEN System. PLoS One 8, e76004PubMedCentralCrossRefPubMedGoogle Scholar
  21. 21.
    Li C, Qi R, Singleterry R et al (2014) Simultaneous gene editing by injection of mRNAs encoding transcription activator-like effector nucleases into mouse zygotes. Mol Cell Biol 34:1649–1658PubMedCentralCrossRefPubMedGoogle Scholar
  22. 22.
    Yasue A, Mitsui SN, Watanabe T et al (2014) Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by the TALEN and CRISPR/Cas systems. Sci Rep 4:5705PubMedCentralCrossRefPubMedGoogle Scholar
  23. 23.
    Sommer D, Peters A, Wirtz T et al (2014) Efficient genome engineering by targeted homologous recombination in mouse embryos using transcription activator-like effector nucleases. Nat Commun 5:3045CrossRefPubMedGoogle Scholar
  24. 24.
    Jones JM, Meisler MH (2014) Modeling human epilepsy by TALEN targeting of mouse sodium channel Scn8a. Genesis 52:141–148CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Institute of Developmental GeneticsHelmholtz Zentrum München, German Research Center for Environmental HealthNeuherbergGermany
  2. 2.Deutsches Zentrum für Neurodegenerative Erkrankungen e. V. (DZNE)MunichGermany
  3. 3.Technische Universität MünchenFreising-WeihenstephanGermany
  4. 4.Max Planck Institute of PsychiatryMunichGermany
  5. 5.Munich Cluster for Systems Neurology (SyNergy)Ludwig-Maximilians-Universität MünchenMunichGermany
  6. 6.Max-Delbrück-Center for Molecular MedicineBerlinGermany
  7. 7.Berlin Institute of HealthBerlinGermany

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