Activation of a diluted E. coli cell-free transcription-translation system within liposomes by hypertonic concentration

Yukino Matsui; Toshiki Akui; Nobuhide Doi; Kei Fujiwara

doi:10.1016/j.xpro.2021.101003

Activation of a diluted E. coli cell-free transcription-translation system within liposomes by hypertonic concentration

Yukino Matsui, Toshiki Akui, Nobuhide Doi, Kei Fujiwara

Department of Biosciences and Informatics

Research output: Contribution to journal › Article › peer-review

1 Citation (Scopus)

Abstract

We present a protocol for activating protein synthesis in liposomes encapsulating a diluted E. coli cell extract-based TX-TL (transcription-translation) system by hypertonic concentration. Protein expression is turned on in the liposome-encapsulated TX-TL system by simple treatment with a concentrated external solution. The expression of sfGFP is demonstrated here, but it can be applied to other proteins. This protocol can be applied to the development of artificial cells utilizing the switch-on mechanism to activate protein expression, responding to the outer environment. For complete details on the use and execution of this protocol, please refer to Akui et al. (2021).

Original language	English
Article number	101003
Journal	STAR Protocols
Volume	2
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2021.101003
Publication status	Published - 2021 Dec 17

Keywords

Biophysics
Biotechnology and bioengineering
Molecular Biology
Protein Biochemistry

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)
Medicine(all)

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10.1016/j.xpro.2021.101003

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title = "Activation of a diluted E. coli cell-free transcription-translation system within liposomes by hypertonic concentration",

abstract = "We present a protocol for activating protein synthesis in liposomes encapsulating a diluted E. coli cell extract-based TX-TL (transcription-translation) system by hypertonic concentration. Protein expression is turned on in the liposome-encapsulated TX-TL system by simple treatment with a concentrated external solution. The expression of sfGFP is demonstrated here, but it can be applied to other proteins. This protocol can be applied to the development of artificial cells utilizing the switch-on mechanism to activate protein expression, responding to the outer environment. For complete details on the use and execution of this protocol, please refer to Akui et al. (2021).",

keywords = "Biophysics, Biotechnology and bioengineering, Molecular Biology, Protein Biochemistry",

author = "Yukino Matsui and Toshiki Akui and Nobuhide Doi and Kei Fujiwara",

note = "Funding Information: We thank Prof. Masahiro Takinoue (Tokyo Institute of Technology) and Prof. Shin-ichiro M. Nomura (Tohoku university) for collaborating on the original work. We thank the financial support by JSPS KAKENHI Grant Number JP15H00826 and JP17H05235 for KF. Funding Information: We thank Prof. Masahiro Takinoue (Tokyo Institute of Technology) and Prof. Shin-ichiro M. Nomura (Tohoku university) for collaborating on the original work. We thank the financial support by JSPS KAKENHI Grant Number JP15H00826 and JP17H05235 for KF. Y.M. T.A. and K.F. established the protocol. T.A. N.D. and K.F. contributed to the original set-up of the experimental system. Y.M. and K.F. wrote the paper. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

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doi = "10.1016/j.xpro.2021.101003",

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AU - Fujiwara, Kei

N1 - Funding Information: We thank Prof. Masahiro Takinoue (Tokyo Institute of Technology) and Prof. Shin-ichiro M. Nomura (Tohoku university) for collaborating on the original work. We thank the financial support by JSPS KAKENHI Grant Number JP15H00826 and JP17H05235 for KF. Funding Information: We thank Prof. Masahiro Takinoue (Tokyo Institute of Technology) and Prof. Shin-ichiro M. Nomura (Tohoku university) for collaborating on the original work. We thank the financial support by JSPS KAKENHI Grant Number JP15H00826 and JP17H05235 for KF. Y.M. T.A. and K.F. established the protocol. T.A. N.D. and K.F. contributed to the original set-up of the experimental system. Y.M. and K.F. wrote the paper. The authors declare no competing interests. Publisher Copyright: © 2021 The Author(s)

PY - 2021/12/17

Y1 - 2021/12/17

N2 - We present a protocol for activating protein synthesis in liposomes encapsulating a diluted E. coli cell extract-based TX-TL (transcription-translation) system by hypertonic concentration. Protein expression is turned on in the liposome-encapsulated TX-TL system by simple treatment with a concentrated external solution. The expression of sfGFP is demonstrated here, but it can be applied to other proteins. This protocol can be applied to the development of artificial cells utilizing the switch-on mechanism to activate protein expression, responding to the outer environment. For complete details on the use and execution of this protocol, please refer to Akui et al. (2021).

AB - We present a protocol for activating protein synthesis in liposomes encapsulating a diluted E. coli cell extract-based TX-TL (transcription-translation) system by hypertonic concentration. Protein expression is turned on in the liposome-encapsulated TX-TL system by simple treatment with a concentrated external solution. The expression of sfGFP is demonstrated here, but it can be applied to other proteins. This protocol can be applied to the development of artificial cells utilizing the switch-on mechanism to activate protein expression, responding to the outer environment. For complete details on the use and execution of this protocol, please refer to Akui et al. (2021).

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KW - Protein Biochemistry

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Activation of a diluted E. coli cell-free transcription-translation system within liposomes by hypertonic concentration

Abstract

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