Single-Copy Insertion of Split-GFP for the Restriction of Germline Expression in Caenorhabditis elegans
Type
ThesisAuthors
Al Johani, Mohammed
Advisors
Frøkjær-Jensen, Christian
Committee members
Adamo, Antonio
Krattinger, Simon G.

Program
BioscienceDate
2018-11Embargo End Date
2019-12-06Permanent link to this record
http://hdl.handle.net/10754/630160
Metadata
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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-4H100ae974a485f413a2113503eed53cd6c53
10.25781/KAUST-4H100