Genome Editing in Mice Using TALE Nucleases

Methods Mol Biol. 2016:1338:229-43. doi: 10.1007/978-1-4939-2932-0_17.

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.

Keywords: Gene targeting; Knock-in; Knockout; Mouse; One-cell embryo; TALEN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics*
  • DNA Breaks, Double-Stranded
  • Endonucleases / genetics*
  • Gene Knockout Techniques / methods*
  • Genome
  • Mice
  • Microinjections
  • Mutation
  • RNA Editing / genetics*
  • RNA, Messenger / genetics

Substances

  • RNA, Messenger
  • Endonucleases