Zinc-finger nucleases: the next generation emerges

Mol Ther. 2008 Jul;16(7):1200-7. doi: 10.1038/mt.2008.114. Epub 2008 Jun 10.

Abstract

Methods of modifying the human genome precisely and efficiently hold great promise for revolutionizing the gene therapy arena. One particularly promising technology is based on the homologous recombination (HR) pathway and is known as gene targeting. Until recently, the low frequency of HR in mammalian cells, and the resulting dependence on selection to identify these rare events, has prevented gene targeting from being applied in a therapeutic context. However, recent advances in generating customized zinc-finger nucleases (ZFNs) that can create a DNA double-strand break (DSB) at preselected sites in the human genome have paved the way for HR-based strategies in gene therapy. By introducing a DSB into a target locus of interest, ZFNs stimulate gene targeting by several orders of magnitude through activation of cellular DNA repair pathways. The capability of this technology to achieve gene conversion frequencies of up to 29% in the absence of selection demonstrates its potential power. In this paper we review recent advances in, and upcoming challenges for, this emerging technology and discuss future experimental work that will be needed to bring ZFNs safely into a clinical setting.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • DNA Breaks, Double-Stranded*
  • Deoxyribonucleases / genetics
  • Deoxyribonucleases / metabolism*
  • Deoxyribonucleases / toxicity
  • Gene Targeting / methods*
  • Genetic Engineering
  • Genetic Therapy / methods*
  • Genome, Human*
  • Humans
  • Mice
  • Zinc Fingers*

Substances

  • Deoxyribonucleases