Gene Editing in One-Cell Embryos by Zinc-Finger and TAL Nucleases

Benedikt Wefers; Melanie Meyer; Svenja Hensler; Sudeepta Panda; Oskar Ortiz; Wolfgang Wurst; Ralf Kühn

doi:10.1002/9780470942390.mo120177

Gene Editing in One-Cell Embryos by Zinc-Finger and TAL Nucleases

Curr Protoc Mouse Biol. 2012 Dec 1;2(4):347-64. doi: 10.1002/9780470942390.mo120177.

Authors

Benedikt Wefers¹, Melanie Meyer^{1

2}, Svenja Hensler^{1

2}, Sudeepta Panda^{1

2}, Oskar Ortiz¹, Wolfgang Wurst^{1

2

3

4}, Ralf Kühn^{1

2}

Affiliations

¹ Helmholtz Center Munich, Institute for Developmental Genetics Munich, Germany.
² Chair for Developmental Genetics, Technische Universität München, Munich, Germany.
³ Max Planck Institute of Psychiatry, Molecular Neurogenetics, Munich, Germany.
⁴ Deutsches Zentrum für Neurodegenerative Erkrankungen, Standort München, Munich, Germany.

PMID: 26069019
DOI: 10.1002/9780470942390.mo120177

Abstract

Gene targeting by sequence-specific nucleases in one-cell embryos provides an expedited mutagenesis approach in rodents. This technology has been recently established to create knockout and knockin mutants through sequence deletion or sequence insertion. This article provides protocols for the preparation and microinjection of nuclease mRNA and targeting vector DNA into fertilized mouse eggs. Furthermore, we provide guidelines for genotyping the desired mouse mutants. Curr. Protoc. Mouse Biol. 2:347-364 © 2012 by John Wiley & Sons, Inc.

Keywords: TAL nuclease; gene targeting; homologous recombination; mouse mutant; pronucleus injection; zinc-finger nuclease.