@article{7be9af41bbfb42abac3ff3261d5487f4,
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)",
year = "2021",
month = dec,
day = "17",
doi = "10.1016/j.xpro.2021.101003",
language = "English",
volume = "2",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
number = "4",
}