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      Targeting DNA Double-Strand Breaks with TAL Effector Nucleases

      Author(s): , , , , , , ,
      Publication date Created:
      Publication date (Electronic):
      Journal: Genetics
      Publisher: Genetics Society of America

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          Abstract

          Engineered nucleases that cleave specific DNA sequences in vivo are valuable reagents for targeted mutagenesis. Here we report a new class of sequence-specific nucleases created by fusing transcription activator-like effectors (TALEs) to the catalytic domain of the FokI endonuclease. Both native and custom TALE-nuclease fusions direct DNA double-strand breaks to specific, targeted sites.

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          Chimeric nucleases stimulate gene targeting in human cells.

          Matthew Porteus, David Baltimore (2003)
          Article 0 comments Cited 207 times – based on 0 reviews     Review now
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            High frequency modification of plant genes using engineered zinc finger nucleases

            Jeffrey A. Townsend, David Wright, Ronnie J. Winfrey … (2009)
            An efficient method for making directed DNA sequence modifications to plant genes (gene targeting) is presently lacking, thereby frustrating efforts to dissect plant gene function and engineer crop plants that better produce the world's burgeoning need for food, fiber and fuel. Zinc finger nucleases (ZFNs) - enzymes engineered to create DNA double strand breaks at specific loci - are potent stimulators of gene targeting,1,2 including at engineered reporter genes in plants.3,4 Here we demonstrate high frequency ZFN-stimulated gene targeting at endogenous plant genes, namely the tobacco acetohydroxyacid synthase (SuRA and SuRB) genes, for which specific mutations are known to confer resistance to imidazolinone and sulfonylurea herbicides.5 Herbicide-resistance mutations were introduced into SuR loci by ZFN-mediated gene targeting at frequencies exceeding 2% of transformed cells for mutations as far as 1.3 kb from the ZFN cleavage site. More than 40% of recombinant plants had modifications in multiple SuR alleles. The observed high frequency of gene targeting indicates that it is now possible to efficiently make targeted sequence changes in endogenous plant genes.
            Article 0 comments Cited 205 times – based on 0 reviews     Review now
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              Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification.

              Morgan Maeder, Stacey Thibodeau-Beganny, Anna Osiak … (2008)
              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 zinc-finger arrays. Here we describe OPEN (Oligomerized Pool ENgineering), a rapid, publicly available strategy for constructing multifinger 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%-50%) at 11 different target sites located within three endogenous human genes (VEGF-A, HoxB13, and CFTR), an endogenous plant gene (tobacco SuRA), and a chromosomally integrated EGFP reporter gene. In summary, OPEN provides an "open-source" 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.
              Article 0 comments Cited 203 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Title: Genetics
                Abbreviated Title: Genetics
                Publisher: Genetics Society of America
                ISSN (Print): 0016-6731
                ISSN (Electronic): 1943-2631
                Publication date Created: October 12 2010
                Publication date Created: October 2010
                Publication date (Print): October 2010
                Publication date (Electronic): July 26 2010
                Volume: 186
                Issue: 2
                Pages: 757-761
                Article
                DOI: 10.1534/genetics.110.120717
                PMC ID: 2942870
                PubMed ID: 20660643
                SO-VID: 2c9724a2-af92-4473-b534-68ebaecc34d5
                Copyright © © 2010
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