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    Single-Copy Insertion of Split-GFP for the Restriction of Germline Expression in Caenorhabditis elegans

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    Name:
    Aljohnani_M_Thesis_Final.pdf
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    Description:
    MS Thesis_Final
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    Type
    Thesis
    Authors
    Al Johani, Mohammed cc
    Advisors
    Frøkjær-Jensen, Christian cc
    Committee members
    Adamo, Antonio cc
    Krattinger, Simon G. cc
    Program
    Bioscience
    KAUST Department
    Biological and Environmental Sciences and Engineering (BESE) Division
    Date
    2018-11
    Embargo End Date
    2019-12-06
    Permanent link to this record
    http://hdl.handle.net/10754/630160
    
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    Access Restrictions
    At the time of archiving, the student author of this thesis opted to temporarily restrict access to it. The full text of this thesis became available to the public after the expiration of the embargo on 2019-12-06.
    Abstract
    Gene regulation in C. elegans germ cells depend on transgenerational chromatin modification and small RNA pathways. Germline silencing mechanisms evolved to repress foreign DNA from compromising the transfer of genetic information to progeny. Effective genetic tools that circumvent the silencing machinery will facilitate studies using this model organism. Specifically, translation of heat-shock inducible transgenes is inhibited in the germline making it challenging to transiently express enzymes to modify the genome. Here, we describe a genetic screen design that can be used to identify pathways that prevent germline expression of heat-shock induced transgenes. We use split-GFP (GFP1-10 and GFP11) to confine a genetic screen to germ cells. Stable transgenic lines with germline expression of single-copy integrated GFP11 were produced using MosSCI. The insertion lines will be used in RNAi or chemical mutagenesis screens for the germline de-repression of GFP1-10 expressed under heat-shock promoters. The screen is likely to identify candidate RNAi or chromatin factors involved in repressing heat-shock expression in the germline, particularly from extrachromosomal arrays. Inducible high-level expression in the germline from extrachromosomal arrays would be a valuable tool for large-scale genome engineering.
    Citation
    Al Johani, M. (2018). Single-Copy Insertion of Split-GFP for the Restriction of Germline Expression in Caenorhabditis elegans. KAUST Research Repository. https://doi.org/10.25781/KAUST-4H100
    DOI
    10.25781/KAUST-4H100
    ae974a485f413a2113503eed53cd6c53
    10.25781/KAUST-4H100
    Scopus Count
    Collections
    Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program; Theses

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