Implementing The Twinkle Factory reagents
Frequently asked questionsNewbee getting started with FAST and splitFAST? Already an established player? Check here the most frequent questions asked about FAST, splitFAST, and CATCHFIRE
Q. Can you recommend a linker between FAST and my POI?
We usually use SGGGSGGGS as a linker. Now, you may want to adapt according to your POI other
Q. Can I implement FAST and splitFAST on fixed cells?
Yes! Despite unpublished, several teams have been using them on fixed cells
Q. Do FAST and splitFAST work at high temperature?
They do! Researchers of TU Wien indeed found pFAST highly functional in Thermoanaerobacter kivui even at 66°C (151°F) growth temperature. They performed measurements both in flow cytometry and with a plate reader photometer at 30°C (Caution, the fluorescence of the assembly FAST-Fluorogen drastically diminishes above 55°C, but reversibly!)
Q. In which anaerobes have FAST and splitFAST been used so far?
Since the initial developments by Prof. Papoutsakis of University of Delaware in 2019, FAST and splitFAST have equipped a variety of anaerobic bacteria:
- Acetobacterium woodii
- Clostridium acetobutylicum
- Clostridium saccharoperbutylacetonicum
- Eubacterium limosum
Interestingly, can one use FAST and splitFAST in usually-non-anaerobic bacteria while exposed to hypoxic conditions:
- in biofilms (E. coli MG1655-F, Bacillus thuringiensis)
- in the gut and in tumors (E. coli Nissle 1917) or in the cystic fibrosis lung (Pseudomonas aeruginosa)
Beyond anaerobic bacteria, FAST and splitFAST were also proven in archaea:
And, finally, in anaerobic protozoa:
Q. Can I use splitFAST to spy on ternary protein assemblies?
Yes, indeed! Check bioRxiv 2023 by Prof. A. Gautier team
Q. Was CATCHFIRE reported since the princeps paper in Nat. Methods in August 2023?
Nope! Give it a try!