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Boolean Logic-responsive FRET Biosensors via Genetically Encoded Autonomous Compilation

Son, H.; Hoye, J. W.; Khawand, M.; Ross, M. L.; Gharios, R.; DeForest, C. A.

2026-09-24 synthetic biology
10.64898/2026.09.24.753966 bioRxiv
Show abstract

Forster resonance energy transfer (FRET) is commonly used to monitor protein-protein interactions in situ. The high spatiotemporal resolution and facile implementation inside complex molecular environments have spearheaded FRET's widespread adoption in biosensing. Despite these advantages, current FRET biosensors are largely restricted to the detection of the presence/absence of individual inputs and are thus unable to sense several multiplexable inputs simultaneously within complex milieu of biological environments. In this work, we introduce a generalizable strategy to construct genetically encoded protein-based FRET biosensors capable of recognizing multiple inputs following Boolean logic-type (YES/OR/AND) operations. These topologically specified FRET sensors powerfully expand the input capacity in sensing protein-protein interactions while providing a user-programmable platform for monitoring heterogeneous biological activities both in vitro and in living cells.

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