Comparison of the efficiency of TIR1 transgenes to provoke auxin induced LAG-1 degradation in Caenorhabditis elegans germline stem cells release_ztjdizmh6bgrtah5z7m2yv43cm

by Jian Chen, Ariz Mohammad, Tim Schedl

Published in microPublication Biology by microPublication Biology.

2020   Volume 2020

Abstract

The auxin inducible degradation (AID) system was first introduced in C. elegans by Dernburg lab and has become a widely-used approach to study tissue-specific and/or temporal aspects of gene function (Zhang et al. 2015; Ashley et al. 2020; Martinez et al. 2020). The AID system utilized a plant derived E3 ubiquitin ligase, TIR1, to specifically degrade proteins fused with the "degron" tag following treatment with the plant growth hormone auxin. To increase the utility of the AID system, the Ward laboratory recently generated an expanded set of TIR1s transgenes, controlled by different tissue-specific promoters (Ashley et al. 2020). Here we aim to compare different germline-expressed TIR1 transgenes for their efficiency in degrading the transcription factor LAG-1, which is C-terminally tagged with degron (Chen et al., 2020). As indicated in Figure 1, these TIR1 transgenes are driven by the following promoters: gld-1p (Zhang et al. 2015), mex-5p (Ashley et al. 2020), sun-1p (Ashley et al. 2020), and pie-1p (Kasimatis et al. 2018), and contain the indicated C-terminal fluorescent proteins and 3' untranslated regions (Fig. 1A).
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