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The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice
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Journal Article
The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice
2020
Overview
Protein knockdown using the auxin-inducible degron (AID) technology is useful to study protein function in living cells because it induces rapid depletion, which makes it possible to observe an immediate phenotype. However, the current AID system has two major drawbacks: leaky degradation and the requirement for a high dose of auxin. These negative features make it difficult to control precisely the expression level of a protein of interest in living cells and to apply this method to mice. Here, we overcome these problems by taking advantage of a bump-and-hole approach to establish the AID version 2 (AID2) system. AID2, which employs an OsTIR1(F74G) mutant and a ligand, 5-Ph-IAA, shows no detectable leaky degradation, requires a 670-times lower ligand concentration, and achieves even quicker degradation than the conventional AID. We demonstrate successful generation of human cell mutants for genes that were previously difficult to deal with, and show that AID2 achieves rapid target depletion not only in yeast and mammalian cells, but also in mice.
Auxin-inducible degron systems can be leaky and require high doses of auxin. Here the authors establish AID2 which uses an OsTIR1 mutant and the ligand 5-Ph-IAA to overcome these problems and establish AID-mediated target depletion in mice.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 64/60
/ 82/80
/ Animals
/ Cell Cycle Proteins - genetics
/ Cell Cycle Proteins - metabolism
/ DNA-Binding Proteins - genetics
/ DNA-Binding Proteins - metabolism
/ Female
/ Humanities and Social Sciences
/ Humans
/ Indoleacetic Acids - pharmacology
/ Ligands
/ Male
/ Mammals
/ Minichromosome Maintenance Proteins - genetics
/ Minichromosome Maintenance Proteins - metabolism
/ Mutants
/ Mutation
/ Nuclear Proteins - metabolism
/ Proteins
/ Recombinant Fusion Proteins - genetics
/ Recombinant Fusion Proteins - metabolism
/ Saccharomyces cerevisiae Proteins - genetics
/ Saccharomyces cerevisiae Proteins - metabolism
/ Science
/ Xenograft Model Antitumor Assays
/ Yeast
