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bioRxiv · 10.1101/2024.01.16.575916

Efficacy of auxin-inducible protein degradation in C. elegans tissues using different auxins and TIR1-expressing strains

Sharma, N.·Marques, F.·Kratsios, P.
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Abstract

The auxin-inducible degradation system has emerged as a powerful tool to deplete proteins of interest in cells and tissues of various model organisms, including C. elegans 2-5. Here, we present a detailed protocol to perform AID-driven spatiotemporal depletion of specific proteins in C. elegans tissues. First, we introduced the AID degron and a fluorescent reporter at two conserved proteins: (a) the transcription factor CFI-1 (human ARID3), which is expressed in the nucleus of multiple C. elegans neurons and head muscle cells 6,7, and (b) the broadly expressed translation initiation factor Y47D3A.21 (human DENR) that localizes in the cytoplasm. Second, we provide a step-by-step guide on how to generate C. elegans strains suitable for AID-mediated protein (CFI-1 and DENR) depletion. Third, we find that the degree of CFI-1 and DENR depletion in C. elegans tissues is comparable upon treatment with either natural auxin (indole-3-acetic acid (IAA) or a water-soluble synthetic auxin analog (K-NAA). Last, we compare the degree of AID-mediated CFI-1 depletion in C. elegans neurons when the transport inhibitor response 1 (TIR1), component of the SCF ubiquitin ligase complex, is provided in neurons or all somatic cells. Altogether, this protocol provides side-by-side comparisons of different auxins and TIR1-expressing lines. Such comparisons may benefit future studies of AID-mediated protein depletion in C. elegans. Graphical abstractImage provided as pdf (together with Figures). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=199 SRC="FIGDIR/small/575916v1_ufig1.gif" ALT="Figure 1"> View larger version (37K): org.highwire.dtl.DTLVardef@adf68eorg.highwire.dtl.DTLVardef@1160cfcorg.highwire.dtl.DTLVardef@aeb593org.highwire.dtl.DTLVardef@12c63e_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIEfficient protein depletion in C. elegans tissues upon treatment with either natural or synthetic auxins. C_LIO_LIPansomatic TIR1 expression leads to efficient depletion of CFI-1 and DENR. C_LIO_LIPanneuronal TIR1 expression leads to neuron-specific, yet variable CFI-1 depletion. C_LIO_LIThe AID system is compatible with fluorescence microscopy, Western blotting and behavioral assays. C_LI

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Sharma, N., Marques, F., Kratsios, P.. 2024-01-20. Efficacy of auxin-inducible protein degradation in C. elegans tissues using different auxins and TIR1-expressing strains. https://doi.org/10.1101/2024.01.16.575916

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