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    RNA-guided Transcriptional Regulation in Plants via dCas9 Chimeric Proteins

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    Hatoon Baazim Thesis.pdf
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    Hatoon Baazim Thesis
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    Type
    Thesis
    Authors
    Baazim, Hatoon
    Advisors
    Mahfouz, Magdy M. cc
    Committee members
    Gehring, Christoph A cc
    Hamdan, Samir cc
    Program
    Bioscience
    KAUST Department
    Biological and Environmental Science and Engineering (BESE) Division
    Date
    2014-05
    Permanent link to this record
    http://hdl.handle.net/10754/316715
    
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    Abstract
    Developing targeted genome regulation approaches holds much promise for accelerating trait discovery and development in agricultural biotechnology. Clustered Regularly Interspaced Palindromic Repeats (CRISPRs)/CRISPR associated (Cas) system provides bacteria and archaea with an adaptive molecular immunity mechanism against invading nucleic acids through phages and conjugative plasmids. The type II CRISPR/Cas system has been adapted for genome editing purposes across a variety of cell types and organisms. Recently, the catalytically inactive Cas9 (dCas9) protein combined with guide RNAs (gRNAs) were used as a DNA-targeting platform to modulate the expression patterns in bacterial, yeast and human cells. Here, we employed this DNA-targeting system for targeted transcriptional regulation in planta by developing chimeric dCas9-based activators and repressors. For example, we fused to the C-terminus of dCas9 with the activation domains of EDLL and TAL effectors, respectively, to generate transcriptional activators, and the SRDX repression domain to generate transcriptional repressor. Our data demonstrate that the dCas9:EDLL and dCas9:TAD activators, guided by gRNAs complementary to promoter elements, induce strong transcriptional activation on episomal targets in plant cells. Moreover, our data suggest that the dCas9:SRDX repressor and the dCas9:EDLL and dCas9:TAD activators are capable of markedly repressing or activating, respectively, the transcription of an endogenous genomic target. Our data indicate that the CRISPR/dCas9:TFs DNA targeting system can be used in plants as a functional genomic tool and for biotechnological applications.
    Citation
    Baazim, H. (2014). RNA-guided Transcriptional Regulation in Plants via dCas9 Chimeric Proteins. KAUST Research Repository. https://doi.org/10.25781/KAUST-356X6
    DOI
    10.25781/KAUST-356X6
    ae974a485f413a2113503eed53cd6c53
    10.25781/KAUST-356X6
    Scopus Count
    Collections
    Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; MS Theses

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