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Article Abstract
Cryptococcus neoformans is the top-ranked W.H.O. fungal priority pathogen, but tools for generating conditional mutations are limited. Auxin-inducible degron systems permit rapid and effective cellular depletion of a tagged protein of interest upon adding a small molecule. These tools are invaluable, particularly for studying essential genes, which may play important roles in pathogen biology. AID2 is one such system that improves on previous strategies. This system achieves greater sensitivity and specificity through an auxin derivative, 5-Ph-IAA, alongside an OsTIR1F74G mutant. We adapted the AID2 system for C. neoformans by codon optimizing OsTIR1F74G and tested its use in multiple scenarios. We demonstrate that the C. neoformans optimized AID2 system enables effective degradation of proteins, including essential proteins, and can be used to help discriminate essential from nonessential genes. This tool enables the study of unexplored parts of the C. neoformans genome.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC12134991 | PMC |
| http://dx.doi.org/10.1093/g3journal/jkaf071 | DOI Listing |
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View Article and Find Full Text PDFArticle Synopsis
- The W.H.O. fungal priority pathogen requires better tools for studying essential genes, and the auxin-inducible degron (AID) system offers a solution for rapidly depleting proteins.
- The AID2 system enhances previous methods by using a specific auxin (5-Ph-IAA) and a mutant version of OsTIR1 to increase sensitivity and effectiveness in protein degradation.
- Researchers optimized the AID2 system for their studies, proving it can effectively target and degrade essential proteins, thus allowing for deeper exploration of the genome's functionality.