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The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice
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 re...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7659001/ https://www.ncbi.nlm.nih.gov/pubmed/33177522 http://dx.doi.org/10.1038/s41467-020-19532-z |
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author | Yesbolatova, Aisha Saito, Yuichiro Kitamoto, Naomi Makino-Itou, Hatsune Ajima, Rieko Nakano, Risako Nakaoka, Hirofumi Fukui, Kosuke Gamo, Kanae Tominari, Yusuke Takeuchi, Haruki Saga, Yumiko Hayashi, Ken-ichiro Kanemaki, Masato T. |
author_facet | Yesbolatova, Aisha Saito, Yuichiro Kitamoto, Naomi Makino-Itou, Hatsune Ajima, Rieko Nakano, Risako Nakaoka, Hirofumi Fukui, Kosuke Gamo, Kanae Tominari, Yusuke Takeuchi, Haruki Saga, Yumiko Hayashi, Ken-ichiro Kanemaki, Masato T. |
author_sort | Yesbolatova, Aisha |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7659001 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-76590012020-11-17 The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice Yesbolatova, Aisha Saito, Yuichiro Kitamoto, Naomi Makino-Itou, Hatsune Ajima, Rieko Nakano, Risako Nakaoka, Hirofumi Fukui, Kosuke Gamo, Kanae Tominari, Yusuke Takeuchi, Haruki Saga, Yumiko Hayashi, Ken-ichiro Kanemaki, Masato T. Nat Commun Article 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. Nature Publishing Group UK 2020-11-11 /pmc/articles/PMC7659001/ /pubmed/33177522 http://dx.doi.org/10.1038/s41467-020-19532-z Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Yesbolatova, Aisha Saito, Yuichiro Kitamoto, Naomi Makino-Itou, Hatsune Ajima, Rieko Nakano, Risako Nakaoka, Hirofumi Fukui, Kosuke Gamo, Kanae Tominari, Yusuke Takeuchi, Haruki Saga, Yumiko Hayashi, Ken-ichiro Kanemaki, Masato T. The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title_full | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title_fullStr | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title_full_unstemmed | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title_short | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
title_sort | auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7659001/ https://www.ncbi.nlm.nih.gov/pubmed/33177522 http://dx.doi.org/10.1038/s41467-020-19532-z |
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