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Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification
Molecular Cell
  • Morgan L. Maeder, Massachusetts General Hospital
  • Stacey Thibodeau-Beganny, Massachusetts General Hospital
  • Anna Osiak, Charité Medical School
  • David A. Wright, Iowa State University
  • Reshma M. Anthony, University of Iowa
  • Magdalena Eichtinger, Massachusetts General Hospital
  • Tao Jiang, Massachusetts General Hospital
  • Jonathan E. Foley, Massachusetts General Hospital
  • Ronnie J. Winfrey, Iowa State University
  • Jeffrey A. Townsend, Iowa State University
  • Erica Unger-Wallace, Iowa State University
  • Jeffry D. Sander, Iowa State University
  • Felix Müller-Lerch, Charité Medical School
  • Fengli Fu, Iowa State University
  • Joseph Pearlberg, Harvard Medical School
  • Carl Göbel, Massachusetts General Hospital
  • Justin P. Dassie, University of Iowa
  • Shondra M. Pruett-Miller, UT Southwestern Medical Center
  • Matthew H. Porteus, UT Southwestern Medical Center
  • Dennis C. Sgroi, Massachusetts General Hospital
  • A. John Iafrate, Harvard Medical School
  • Drena Dobbs, Iowa State University
  • Paul B. McCray, Jr., University of Iowa
  • Toni Cathomen, Charité Medical School
  • Daniel F. Voytas, Iowa State University
  • J. Keith Joung, Massachusetts General Hospital
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Publication Version
Accepted Manuscript
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Summary—Custom-made zinc-finger nucleases (ZFNs) can induce targeted genome modifications with high efficiency in cell types including Drosophila, C. elegans, plants, and humans. A bottleneck in the application of ZFN technology has been the generation of highly specific engineered zincfinger arrays. Here we describe OPEN (Oligomerized Pool ENgineering), a rapid, publicly available strategy for constructing multi-finger arrays, which we show is more effective than the previously published modular assembly method. We used OPEN to construct 37 highly active ZFN pairs which induced targeted alterations with high efficiencies (1 to 50%) at 11 different target sites located within three endogenous human genes (VEGF-A, HoxB13, CFTR), an endogenous plant gene (tobacco SuRA), and a chromosomally-integrated EGFP reporter gene. In summary, OPEN provides an “opensource” method for rapidly engineering highly active zinc-finger arrays, thereby enabling broader practice, development, and application of ZFN technology for biological research and gene therapy.

This is a manuscript of an article from Molecular Cell 31 (2008): 294, doi: 10.1016/j.molcel.2008.06.016. Posted with permission.

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Elsevier Inc.
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Citation Information
Morgan L. Maeder, Stacey Thibodeau-Beganny, Anna Osiak, David A. Wright, et al.. "Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification" Molecular Cell Vol. 31 Iss. 2 (2008) p. 294 - 301
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