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Resources » Paper

Li, Chengyin et al. (2021) International Worm Meeting "Detection of induced gene repression using an in vivo protein recruitment system"

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    Status:
    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00063431

    Li, Chengyin, & Saltzman, Arneet L. (2021). Detection of induced gene repression using an in vivo protein recruitment system presented in International Worm Meeting. Unpublished information; cite only with author permission.

    Gene repression and heterochromatinization are associated with binding of various chromatin regulatory proteins and complexes. Ectopic recruitment of a protein of interest to a reporter locus is a powerful approach used widely in mammalian cells, Drosophila, and fungi to study transcriptional repression and heterochromatin formation. To develop an in vivo tethering system for investigating gene repression in C. elegans, we adapted components of the cGAL GAL4-UAS system (Wang et al. 2017 Nature Methods). The GAL4 DNA-binding domain (DBD) without the activation domain was fused to a protein of interest and recruited to a 15xUAS upstream of a single copy GFP reporter. We confirmed successful tethering of Flag-tagged GAL4 fusion proteins at the UAS by ChIP-qPCR. To establish a positive control to drive gene repression in our system, we constructed GAL4 DBD fused with HPL-2, a homologue of heterochromatin protein 1, and MIG-32, a predicted homologue of a polycomb repressive complex 1 subunit. Fluorescence quantification indicated a significant reduction of GFP intensity upon HPL-2 or MIG-32 recruitment at 1.6-fold and 1.9-fold, respectively. With further optimization, our recruitment system can be utilized to investigate the initiation of gene repression by repressive complex subunits and other heterochromatin-associated proteins in C. elegans.

    Affiliation:
    - Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada


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