Genetically encoded tools for tracking metabolites in live cells

Area of Science:

• Biotechnology
• Molecular Biology
• Cellular Imaging

Background:

• Metabolite biosensors allow real-time, in situ measurement of metabolites within living systems.
• Fully genetically encoded metabolite biosensors (fGEMBs) utilize fluorescent proteins linked to ligand-binding domains to report metabolite presence.
• These sensors are expressed and quantified using standard molecular biology and imaging techniques.

Purpose of the Study:

• To review the current state and emerging strategies in metabolite biosensor development.
• To highlight advancements in engineering fGEMBs and the rise of chemigenetic and nucleic acid-based sensors.
• To discuss the challenges and future directions in creating novel metabolite sensors.

Main Methods:

• Genetically encoding fluorescent proteins fused to ligand-binding domains for metabolite detection.
• Utilizing fluorimetry, fluorescence microscopy, and flow cytometry for quantifying sensor output.
• Engineering and optimizing biosensors for specific metabolites and cellular environments.

Main Results:

• fGEMBs offer a powerful method for visualizing metabolite dynamics within cells.
• Emerging strategies are improving the throughput and design of fGEMBs.
• Chemigenetic metabolite biosensors (cGEMBs) and de novo ligand-binding domain design are expanding sensor capabilities.

Conclusions:

• The field of metabolite biosensors has rapidly expanded due to demand for in situ cellular analysis.
• Continued engineering efforts and novel design approaches promise improved and more versatile metabolite sensors.
• Future developments may shift towards de novo designed ligand-binding domains, moving beyond nature's existing repertoire.