Campus Units
Genetics, Development and Cell Biology, Bioinformatics and Computational Biology
Document Type
Article
Publication Version
Accepted Manuscript
Publication Date
2008
Journal or Book Title
Molecular Cell
Volume
31
Issue
2
First Page
294
Last Page
301
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
Recommended Citation
Maeder, Morgan L.; Thibodeau-Beganny, Stacey; Osiak, Anna; Wright, David A.; Anthony, Reshma M.; Eichtinger, Magdalena; Jiang, Tao; Foley, Jonathan E.; Winfrey, Ronnie J.; Townsend, Jeffrey A.; Unger-Wallace, Erica; Sander, Jeffry D.; Müller-Lerch, Felix; Fu, Fengli; Pearlberg, Joseph; Göbel, Carl; Dassie, Justin P.; Pruett-Miller, Shondra M.; Porteus, Matthew H.; Sgroi, Dennis C.; Iafrate, A. John; Dobbs, Drena; McCray, Paul B. Jr.; Cathomen, Toni; Voytas, Daniel F.; and Joung, J. Keith, "Rapid “Open-Source” Engineering of Customized Zinc-Finger Nucleases for Highly Efficient Gene Modification" (2008). Genetics, Development and Cell Biology Publications. 115.
https://lib.dr.iastate.edu/gdcb_las_pubs/115
Included in
Bioinformatics Commons, Cell and Developmental Biology Commons, Computational Biology Commons, Genetics Commons
Comments
This is a manuscript of an article from Molecular Cell 31 (2008): 294, doi: 10.1016/j.molcel.2008.06.016. Posted with permission.