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Keywords:

  • gene targeting;
  • homologous recombination;
  • zinc finger nucleases;
  • TALE nucleases;
  • morpholino oligonucleotides;
  • RNAi;
  • random mutagenesis;
  • reverse genetics

Abstract

Gene targeting in mice, first reported 25 years ago, has led to monumental advances in the understanding of basic biology and human disease. The ability to employ a similarly straightforward method for gene manipulation in other experimental organisms would make their already significant contributions all the more powerful. Here, we briefly outline the strengths and weaknesses of reverse genetics techniques in non-murine model organisms, ending with a more detailed description of two that promise to bring targeted gene modification to the masses: zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Dana Caroll, a forefather of zinc finger technology, and Bo Zhang, among the first to introduce TALEN-targeted mutagenesis to zebrafish, discuss their experience with these techniques, and speculate about the future of the field. Developmental Dynamics 240:2688–2696, 2011. © 2011 Wiley Periodicals, Inc.