Article
Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification
Molecular Cell
Document Type
Article
Disciplines
Publication Version
Accepted Manuscript
Publication Date
1-1-2008
DOI
10.1016/j.molcel.2008.06.016
Abstract
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.
Copyright Owner
Elsevier Inc.
Copyright Date
2008
Language
en
File Format
application/pdf
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 Available at: http://works.bepress.com/drena-dobbs/44/
This is a manuscript of an article from Molecular Cell 31 (2008): 294, doi: 10.1016/j.molcel.2008.06.016. Posted with permission.