Custom-designed zinc finger nucleases: What is next?

J. Wu, K. Kandavelou, S. Chandrasegaran

Research output: Contribution to journalReview articlepeer-review

Abstract

Custom-designed zinc finger nucleases (ZFNs) - proteins designed to cut at specific DNA sequences - combine the non-specific cleavage domain (N) of Fok I restriction endonuclease with zinc finger proteins (ZFPs). Because the recognition specificities of the ZFPs can be easily manipulated experimentally, ZFNs offer a general way to deliver a targeted site-specific double-strand break (DSB) to the genome. They have become powerful tools for enhancing gene targeting - the process of replacing a gene within a genome of cells via homologous recombination (HR) - by several orders of magnitude. ZFN-mediated gene targeting thus confers molecular biologists with the ability to site-specifically and permanently alter not only plant and mammalian genomes but also many other organisms by stimulating HR via a targeted genomic DSB. Site-specific engineering of the plant and mammalian genome in cells so far has been hindered by the low frequency of HR. In ZFN-mediated gene targeting, this is circumvented by using designer ZFNs to cut at the desired chromosomal locus inside the cells. The DNA break is then patched up using the new investigator-provided genetic information and the cells' own repair machinery. The accuracy and high efficiency of the HR process combined with the ability to design ZFNs that target most DNA sequences (if not all) makes ZFN technology not only a powerful research tool for site-specific manipulation of the plant and mammalian genomes, but also potentially for human therapeutics in the future, in particular for targeted engineering of the human genome of clinically transplantable stem cells.

Original languageEnglish (US)
Pages (from-to)2933-2944
Number of pages12
JournalCellular and Molecular Life Sciences
Volume64
Issue number22
DOIs
StatePublished - Nov 2007

Keywords

  • Gene correction
  • Gene targeting
  • Genome engineering
  • Homologous recombination
  • Non-homologous end-joining
  • Site-specific modification
  • Targeted mutagenesis
  • Zinc finger nucleases

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Cellular and Molecular Neuroscience
  • Cell Biology

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