Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene ModificationMolecular Cell
Publication VersionAccepted Manuscript
AbstractSummary—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 OwnerElsevier Inc.
Citation InformationMorgan 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/