The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice PMID: 33177522

ADC Targets in Oncology

2026-04-10

The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice

Nat Commun. 2020 Nov 11;11(1):5701. doi: 10.1038/s41467-020-19532-z.

Aisha Yesbolatova ¹ ² , Yuichiro Saito ¹ , Naomi Kitamoto ¹ ³ , Hatsune Makino-Itou ⁴ , Rieko Ajima ² ⁴ , Risako Nakano ⁵ , Hirofumi Nakaoka ² ⁶ ⁷ , Kosuke Fukui ⁸ , Kanae Gamo ³ , Yusuke Tominari ³ , Haruki Takeuchi ⁵ ⁹ , Yumiko Saga ² ⁴ ¹⁰ , Ken-Ichiro Hayashi ⁸ , Masato T Kanemaki ¹¹ ¹²

Affiliations

1. Department of Chromosome Science, National Institute of Genetics, Research Organization of Information and Systems (ROIS), Yata 1111, Mishima, Shizuoka, 411-8540, Japan.
2. Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Yata 1111, Mishima, Shizuoka, 411-8540, Japan.
3. FIMECS, Inc., Muraoka-Higashi 2-26-1, Fujisawa, Kanagawa, 251-0012, Japan.
4. Department of Gene Function and Phenomics, National Institute of Genetics, ROIS, Yata 1111, Mishima, Shizuoka, 411-8540, Japan.
5. Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo, Tokyo, 113-0033, Japan.
6. Department of Genomics and Evolutionary Biology, National Institute of Genetics, ROIS, Yata 1111, Mishima, Shizuoka, 411-8540, Japan.
7. Department of Cancer Genome Research, Sasaki Institute, Sasaki Foundation, Kandasurugadai 2-2, Chiyoda-ku, Tokyo, 101-0062, Japan.
8. Department of Biochemistry, Okayama University of Science, Ridai-cho 1-1, Okayama, 700-0005, Japan.
9. Social Cooperation Program of Evolutional Chemical Safety Assessment System, LECSAS, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo, Tokyo, 113-0033, Japan.
10. Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, 113-0033, Japan.
11. Department of Chromosome Science, National Institute of Genetics, Research Organization of Information and Systems (ROIS), Yata 1111, Mishima, Shizuoka, 411-8540, Japan. [email protected].
12. Department of Genetics, The Graduate University for Advanced Studies (SOKENDAI), Yata 1111, Mishima, Shizuoka, 411-8540, Japan. [email protected].

Abstract

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.