Switching ON of Transcription-Translation System Using GUV Fusion by Co-supplementation of Calcium with Long-Chain Polyethylene Glycol

Yusuke Uwaguchi, Kei Fujiwara, Nobuhide Doi

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Giant unilamellar vesicles (GUVs) have been used as a material for bottom-up synthetic biology. However, due to the semi-permeability of the membrane, the need for methods to fuse GUVs has increased. To this aim, methods that are simple and show low leakage during fusion are important. In this study, we report a method of GUV fusion by a divalent cation (Ca2+) enhanced with a long chain polyethylene glycol (PEG20k). The methods showed significant GUV fusion without leakage of internal components of GUVs and maintained cell-free transcription-translation ability inside the GUVs without external supplementation of macromolecules. We demonstrate that the Ca-PEG method can be applied for switching ON of transcription-translation in GUVs in a fusion-dependent manner. The method developed here can be applied to extend bottom-up synthetic biology and molecular robotics that use GUVs as a chassis.

Original languageEnglish
Pages (from-to)2319-2324
Number of pages6
JournalChemBioChem
Volume22
Issue number13
DOIs
Publication statusPublished - 2021 Jul 1

Keywords

  • artificial cells
  • bottom-up synthetic biology
  • in vitro transcription-translation
  • liposome fusion
  • liposomes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

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