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    Activities and specificities of homodimeric TALENs in Saccharomyces cerevisiae

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    Type
    Article
    Authors
    Aouida, Mustapha cc
    Piatek, Marek J. cc
    Bangarusamy, Dhinoth Kumar
    Mahfouz, Magdy M. cc
    KAUST Department
    Biological and Environmental Science and Engineering (BESE) Division
    Bioscience Core Lab
    Bioscience Program
    Center for Desert Agriculture
    Laboratory for Genome Engineering
    Plant Science
    Plant Science Program
    Date
    2013-10-01
    Online Publication Date
    2013-10-01
    Print Publication Date
    2014-05
    Permanent link to this record
    http://hdl.handle.net/10754/575585
    
    Metadata
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    Abstract
    The development of highly efficient genome engineering reagents is of paramount importance to launch the next wave of biotechnology. TAL effectors have been developed as an adaptable DNA binding scaffold that can be engineered to bind to any user-defined sequence. Thus, TAL-based DNA binding modules have been used to generate chimeric proteins for a variety of targeted genome modifications across eukaryotic species. For example, TAL effectors fused to the catalytic domain of FokI endonuclease (TALENs) were used to generate site-specific double strand breaks (DSBs), the repair of which can be harnessed to dictate user-desired, genome-editing outcomes. To cleave DNA, FokI endonuclease must dimerize which can be achieved using a pair of TALENs that bind to the DNA targeted in a tail-to-tail orientation with proper spacing allowing the dimer formation. Because TALENs binding to DNA are dependent on their repeat sequences and nucleotides binding specificities, homodimers and heterodimers binding can be formed. In the present study, we used several TALEN monomers with increased repeats binding degeneracy to allow homodimer formation at increased number of genomic loci. We assessed their binding specificities and genome modification activities. Our results indicate that homodimeric TALENs could be used to modify the yeast genome in a site-specific manner and their binding to the promoter regions might modulate the expression of target genes. Taken together, our data indicate that homodimeric TALENs could be used to achieve different engineering possibilities of biotechnological applications and that their transcriptional modulations need to be considered when analyzing their phenotypic effects. © 2013 Springer-Verlag.
    Citation
    Aouida, M., Piatek, M. J., Bangarusamy, D. K., & Mahfouz, M. M. (2013). Activities and specificities of homodimeric TALENs in Saccharomyces cerevisiae. Current Genetics, 60(2), 61–74. doi:10.1007/s00294-013-0412-z
    Sponsors
    We thank the Bioscience Core Facility and Optical, Imaging Core Lab King Abdullah University of Science and Technology KAUST for technical assistance. We also thank Nina Fedoroff and the members of the genome engineering group at KAUST for their helpful discussions and technical assistance throughout the preparation of the manuscript. No conflict of interest declared. This research is funded from the Center for Desert Agriculture and genome engineering group baseline funding.
    Publisher
    Springer Nature
    Journal
    Current Genetics
    DOI
    10.1007/s00294-013-0412-z
    PubMed ID
    24081604
    ae974a485f413a2113503eed53cd6c53
    10.1007/s00294-013-0412-z
    Scopus Count
    Collections
    Articles; Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; Plant Science Program; Bioscience Core Lab; Center for Desert Agriculture

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