Protocol for auxin-inducible protein degradation in C. elegans using different auxins and TIR1-expressing strains
Description
The 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-regulated re-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.
For complete details on the use and execution of this protocol, please refer to Li et al.
Data availability
All data generated for or analyzed in this study are contained in the manuscript and supporting files. Microscopy data will be shared by the lead contact upon request.
Any additional information required to analyze or reanalyze data in this manuscript is also available upon request.
No code was generated to analyze the data.
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Protocol-for-auxin-inducible-protein-degradation.pdf
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Additional details
Identifiers
- DOI
- 10.1016/j.xpro.2024.103133
- Other
- oai:uchicago.tind.io:12703
Funding
- Safadi Program of Excellence in Clinical and Translational Neuroscience
- Pilot Grant Award
- Swiss National Science Foundation (SNSF)
- Postdoctoral fellowship
- National Institutes of Health
- R21 NS108505
- National Institutes of Health
- R01 NS118078