Synthetic biology with an all E. coli TXTL system: Quantitative characterization of regulatory elements and gene circuits

Ryan Marshall, Vincent Noireaux

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Over the past decade, a new generation of cell-free transcription-translation (TXTL) systems has been devised for emerging multidisciplinary applications. The DNA-dependent in vitro protein synthesis technology has been developed to tackle applications in synthetic biology, biological and chemical engineering, as well as quantitative disciplines such as biophysics. In addition to being convenient at the biosafety level, the new TXTL platforms are user-friendly; more affordable; more versatile at the level of transcription, with a TX repertoire covering hundreds of parts; and more powerful, with protein production reaching a few mg/mL in batch and continuous modes. As a consequence, TXTL is rising up as a popular research tool and is used by a growing research community. While TXTL is proving reliable for an increasing number of applications, it is important to gain appropriate TXTL skills, especially for quantitative applications. TXTL has become particularly useful to rapidly prototype genetic devices, from single regulatory elements to elementary circuit motifs. In this chapter, we describe the basic procedures to develop appropriate TXTL practices for the characterization of such genetic parts. We use an all E. coli TXTL system developed in our lab, now commercialized by Arbor Biosciences under the name myTXTL.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages61-93
Number of pages33
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1772
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
This material is based upon work supported by the Defense Advanced Research Projects Agency (contract HR0011-16-C-01-34) and the Office of Naval Research (award N00014-13-1-0074).

Keywords

  • CRISPR
  • E. coli
  • Gene circuits
  • Prototyping
  • Quantitative biology
  • Regulatory elements
  • TXTL (cell-free transcription-translation)

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