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Synthetic protein condensates that recruit and release protein activity in living cells

View ORCID ProfileTatsuyuki Yoshii, Masaru Yoshikawa, Masahiro Ikuta, View ORCID ProfileShinya Tsukiji
doi: https://doi.org/10.1101/2020.11.09.375766
Tatsuyuki Yoshii
1Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan
2PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
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Masaru Yoshikawa
3Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan
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Masahiro Ikuta
1Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan
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Shinya Tsukiji
1Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan
3Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan
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  • For correspondence: stsukiji@nitech.ac.jp
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Abstract

Compartmentation of proteins into biomolecular condensates or membraneless organelles formed by phase separation is an emerging principle for the regulation of cellular processes. Creating synthetic condensates that accommodate specific intracellular proteins on demand would have various applications in chemical biology, cell engineering and synthetic biology. Here, we report the construction of synthetic protein condensates capable of recruiting and/or releasing proteins of interest in living mammalian cells in response to a small molecule or light. We first present chemogenetic protein-recruiting and -releasing condensates, which rapidly inhibited and activated signaling proteins, respectively. An optogenetic condensate system was successfully constructed that enables reversible release and sequestration of protein activity using light. This proof-of-principle work provides a new platform for chemogenetic and optogenetic control of protein activity in mammalian cells and represents a step towards tailor-made engineering of synthetic protein condensates with various functionalities.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted November 10, 2020.
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Synthetic protein condensates that recruit and release protein activity in living cells
Tatsuyuki Yoshii, Masaru Yoshikawa, Masahiro Ikuta, Shinya Tsukiji
bioRxiv 2020.11.09.375766; doi: https://doi.org/10.1101/2020.11.09.375766
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Synthetic protein condensates that recruit and release protein activity in living cells
Tatsuyuki Yoshii, Masaru Yoshikawa, Masahiro Ikuta, Shinya Tsukiji
bioRxiv 2020.11.09.375766; doi: https://doi.org/10.1101/2020.11.09.375766

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