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Adaptable, Turn-On Monobody (ATOM) Fluorescent Biosensors for Multiplexed Detection in Cells
Biorxiv : the Preprint Server for Biology
|April 10, 2023
Summary
Researchers developed adaptable, turn-on monobody (ATOM) biosensors for detecting specific proteins in live cells. These single-molecule fluorescent protein biosensors offer high specificity and rapid adaptation for diverse biological targets.
Area of Science:
- Molecular Biology
- Biophysics
- Biochemistry
Background:
- Developing adaptable biosensors is crucial for molecular detection.
- Fluorescent protein-based platforms offer potential for single-molecule sensing.
- Existing methods face challenges in target specificity and ease of adaptation.
Approach:
- Created adaptable, turn-on monobody (ATOM) biosensors by inserting circularly permuted monobodies into fluorescent proteins.
- Engineered sensors targeting WDR5, SH2, and hRAS proteins, localizing them to nucleus, cytoplasm, and plasma membrane.
- Demonstrated multiplexed imaging in live human cells using cyan, yellow, and red ATOM sensors.
Key Points:
- ATOM biosensors exhibit high specificity in detecting intracellular and organelle-targeted proteins.
- Ligand binding triggers fluorescence activation via chromophore maturation, with >20-fold turn-on in cells and up to 100-fold in vitro.
- The sensing mechanism was validated across different fluorescent protein lineages (jellyfish and anemone).
Conclusions:
- ATOM biosensors provide a versatile, single-molecule platform for detecting diverse targets.
- The modular design allows for rapid generation of new biosensors with varied specificities and colors.
- This technology advances live-cell imaging and molecular sensing capabilities.
