Protocol for auxin-inducible protein degradation in C. elegans using different auxins and TIR1-expressing strains

Protocol for auxin-inducible protein degradation in C. elegans using different auxins and TIR1-expressing strainsThe auxin-inducible degron (AID) system is a powerful tool to deplete proteins in vivo.Here, we present a protocol for AID-mediated depletion of two proteins (CFI-1/AT-rich interaction domain 3 [ARID3] and Y47D3A.21/density-regulatedre-initiation and release factor [DENR]) in C. elegans tissues using different auxins and transport inhibitor response 1 (TIR1)-expressing strains.We describe steps for genetic crossing, sample preparation, fluorescent microscopy, and treatment with either natural (indole-3-acetic acid [IAA]) or synthetic (1-naphthaleneacetic acid, potassium salt [K-NAA]) auxins.We then detail procedures for comparing the degree of CFI-1 depletion in C. elegans neurons upon panneuronal or pansomatic TIR1 expression.