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.

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.

Copyright Owner

Elsevier Inc.

Language

en

File Format

application/pdf

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