TXTL-based approach to synthetic cells

Jonathan Garamella, David Garenne, Vincent Noireaux

Research output: Chapter in Book/Report/Conference proceedingChapter

10 Scopus citations

Abstract

Cell-free transcription–translation (TXTL) has recently emerged as a versatile technology to engineer biological systems. In this chapter, we show how an all E. coli TXTL system can be used to build synthetic cell prototypes. We describe methods to encapsulate TXTL reactions in cell-sized liposomes, with an emphasis on the composition of the external solution and lipid bilayer. Cell-free expression is quantitatively described in bulk reactions and liposomes for three proteins: the soluble reporter protein eGFP, the membrane proteins alpha-hemolysin (AH) from Staphylococcus aureus, and the mechanosensitive channel of large conductance (MscL) from E. coli.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages217-239
Number of pages23
Volume617
DOIs
StatePublished - 2019

Publication series

NameMethods in enzymology
PublisherAcademic Press Inc.
ISSN (Print)0076-6879

Bibliographical note

Funding Information:
This material is based upon work supported by the Human Frontier Science Program (research grant RGP0037/2015), the NSF grant MCB-1613677, and the US Israel Binational Science Foundation.

Funding Information:
This material is based upon work supported by the Human Frontier Science Program (research grant RGP0037/2015), the NSF grant MCB-1613677, and the US Israel Binational Science Foundation., The Noireaux laboratory receives research funds from Arbor Biosciences, a distributor of the myTXTL cell-free protein synthesis kit.

Funding Information:
This material is based upon work supported by the Human Frontier Science Program (research grant RGP0037/2015), the NSF grant MCB-1613677, and the US Israel Binational Science Foundation. The Noireaux laboratory receives research funds from Arbor Biosciences, a distributor of the myTXTL cell-free protein synthesis kit.

Publisher Copyright:
© 2019 Elsevier Inc.

Keywords

  • Bioengineering
  • Cell-free transcription–translation
  • Gene circuits
  • Liposomes
  • Membrane proteins
  • Synthetic biology
  • Synthetic cell
  • Protein Biosynthesis
  • Escherichia coli/genetics
  • Hemolysin Proteins/genetics
  • Genetic Engineering/methods
  • Bacterial Toxins/genetics
  • Green Fluorescent Proteins/genetics
  • Ion Channels/genetics
  • Escherichia coli Proteins/genetics
  • Transcription, Genetic
  • Liposomes/metabolism
  • Staphylococcus aureus/genetics
  • Artificial Cells/metabolism

PubMed: MeSH publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Journal Article

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