Towards a new standard with FAST in archaea imaging! A team led by Silvan Scheller of Aalto University has just reported implementing FAST in Methanococcus maripaludis and Methanosarcina acetivorans in bioRxiv 2022. Only a few weeks ago, had Kyle Costa reported FAST in archaea for the first time ever.
Crucial in the global carbon cycle, methane-producing archaea have a high potential for biofuel production and carbon dioxide capture. They also candidate for high-end electrochemical systems. Of those, Methanococcus maripaludis and Methanosarcina acetivorans are well-characterized methanogenic model organisms. Besides they can be genetically engineered with a variety of molecular tools. However, their anaerobic lifestyle and autofluorescence restrict the use of common fluorescent reporter proteins (e.g., GFP). Those indeed require oxygen for chromophore maturation.
Here, the authors have employed tdFAST, the tandem version of FAST of The Twinkle Factory. tdFAST is indeed highly fluorescent in the presence of the cell-permeable fluorescent ligand tfCoral. tdFAST2 expression in M. acetivorans and M. maripaludis was found not cytotoxic. And tdFAST2:tfCoral fluorescence can be clearly distinguished from the autofluorescence. In flow cytometry experiments, mixed methanogen cultures can be clearly distinguished which allows high-throughput investigations of dynamics within single and mixed cultures.
As a result, these findings are expected to accelerate the exploitation of the methanogens’ biotechnological potential.
The Twinkle Factory offers commercial fluorogens for FAST and splitFAST. tfFox-NP, a bright non-permeant fluorogen, and tfDarth, a “dark” chromophore, have recently completed the product range.
More reading on FAST in archaea imaging
- J. Bacteriol. 2022 – The Fluorescence-Activating and Absorption-Shifting Tag (FAST) Enables Live-Cell Fluorescence Imaging of Methanococcus maripaludis – University of Minnesota