Genome Editing in Rats Using TALE Nucleases

Methods Mol Biol. 2016;1338:245-59. doi: 10.1007/978-1-4939-2932-0_18.

Abstract

The rat is an important animal model to understand gene function and model human diseases. Since recent years, the development of gene-specific nucleases has become important for generating new rat models of human diseases, to analyze the role of genes and to generate human antibodies. Transcription activator-like (TALE) nucleases efficiently create gene-specific knockout rats and lead to the possibility of gene targeting by homology-directed recombination (HDR) and generating knock-in rats. We describe a detailed protocol for generating knockout and knock-in rats via microinjection of TALE nucleases into fertilized eggs. This technology is an efficient, cost- and time-effective method for creating new rat models.

Keywords: Gene editing; Genetic engineering; Genomics; Homology-directed repair; Hprt; Knock-in; Knockout; Nonhomologous end joining; Rat model; Rosa26; TALE nucleases; TALEN; Targeted integration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics*
  • Endonucleases / genetics*
  • Gene Knockout Techniques / methods*
  • Genome
  • Homologous Recombination / genetics
  • Humans
  • RNA, Messenger / genetics
  • Rats
  • Trans-Activators / genetics

Substances

  • RNA, Messenger
  • Trans-Activators
  • Endonucleases