greenFAST and redFAST in anaerobes

Jan 31, 2022 | Anaerobe labeling, FAST and splitFAST

greenFAST and redFAST open the door for multicolor approaches to investigate cell dynamics, gene expression, or protein localization in anaerobes.  This is the conclusion of Maximilian Flaiz et al. in this ACS Synth. Biol. 2022 paper.  Indeed, researchers of Prof. Dr. P. Dürre‘s AG at Ulm University have just disclosed the first-ever usage of greenFAST and redFAST in microbiology.  As a result, they continue building up new genetic tools for acetogenic and solventogenic bacteria studies.

greenFAST and redFAST in anaerobes - © 2022 The Authors

greenFAST and redFAST in anaerobes – © 2022 The Authors

Anaerobic bacteria are expected to be pivotal in the future circular-carbon world.  Indeed, several anaerobes are already genetically accessible.  Various molecular tools are available for metabolic engineering.  Still, the use of bright fluorescent reporters, commonly used in molecular biological approaches, is limited under anaerobic conditions.  This is what FAST of The Twinkle Factory is good at!  In this paper, the authors add greenFAST and redFAST to the genetic toolbox of solventogenic and acetogenic bacteria.  Namely they introduce them into Clostridium saccharoperbutylacetonicum for multicolor approaches. These include the investigation of the co-culture dynamics of metabolically engineered strains. Moreover, they establish a tightly regulated inducible two-plasmid system and use greenFAST and redFAST to track the coexistence and interaction of both plasmids.

Prof. Arnaud Gautier had initially disclosed greenFAST and redFAST for orthogonal fluorescent labeling in Nat. Chem. Biol. 2020.

The Twinkle Factory offers commercial fluorogens specific for both greenFAST, i.e., tfLime, and redFAST, i.e., tfCoral.

 

More reading on strain engineering with FAST

  • ACS Synth. Biol. 2022 – Establishment of Green- and Red-Fluorescent Reporter Proteins Based on the Fluorescence-Activating and Absorption-Shifting Tag for Use in Acetogenic and Solventogenic Anaerobes
  • Nat. Commun. 2021 – Auxin-mediated protein depletion for metabolic engineering in terpene-producing yeast
  • Biotechnol. Biofuels 2021 – Production of the Biocommodities Butanol and Acetone from Methanol with Fluorescent FAST-tagged Proteins using Metabolically Engineered Strains of Eubacterium Limosum
  • mBio 2020 – Interspecies Microbial Fusion and Large-Scale Exchange of Cytoplasmic Proteins and RNA in a Syntrophic Clostridium Coculture
  • Appl. Environ. Microbiol. 2019 – A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)