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

Aisha Yesbolatova, Yuichiro Saito, Naomi Kitamoto, Hatsune Makino-Itou, Rieko Ajima, Risako Nakano, Hirofumi Nakaoka, Kosuke Fukui, Kanae Gamo, Yusuke Tominari, Haruki Takeuchi, Yumiko Saga, Ken-ichiro Hayashi and Masato T. Kanemaki ()
Additional contact information
Aisha Yesbolatova: National Institute of Genetics, Research Organization of Information and Systems (ROIS)
Yuichiro Saito: National Institute of Genetics, Research Organization of Information and Systems (ROIS)
Naomi Kitamoto: National Institute of Genetics, Research Organization of Information and Systems (ROIS)
Hatsune Makino-Itou: National Institute of Genetics, ROIS
Rieko Ajima: The Graduate University for Advanced Studies (SOKENDAI)
Risako Nakano: University of Tokyo
Hirofumi Nakaoka: The Graduate University for Advanced Studies (SOKENDAI)
Kosuke Fukui: Okayama University of Science
Kanae Gamo: FIMECS, Inc.
Yusuke Tominari: FIMECS, Inc.
Haruki Takeuchi: University of Tokyo
Yumiko Saga: The Graduate University for Advanced Studies (SOKENDAI)
Ken-ichiro Hayashi: Okayama University of Science
Masato T. Kanemaki: National Institute of Genetics, Research Organization of Information and Systems (ROIS)

Nature Communications, 2020, vol. 11, issue 1, 1-13

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-19532-z

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