Modular Extracellular Sensor Architecture for Orthogonal Ligand Detection
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Solution Overview
Problem
Existing biosensor technologies, such as the TANGO system, rely on native mechanisms and interactions, making them inefficient and prone to interference from endogenous signaling machinery, limiting their adaptability and ability to detect ligands for which no natural receptor exists.
Innovation Solution
The Modular Extracellular Sensor Architecture (MESA) decouples biosensor systems from endogenous signaling, allowing for orthogonal sensing by incorporating a ligand-binding domain, transmembrane domain, protease domain, and functional domain, enabling detection of ligands without native receptor dependency and facilitating engineering of receptors to recognize novel ligands.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If native mechanisms and interactions are used (TANGO system), then the system can detect ligand-protein interactions, but it is prone to interference from endogenous signaling machinery and has limited adaptability
Solution Approach 1:
The biosensor system is divided into separate functional modules: a ligand-binding domain (extracellular), a transmembrane domain, and a functional domain (intracellular). This segmentation allows each module to be independently optimized and combined, enabling the system to function across different cell types without interference from endogenous signaling pathways.
Solution Approach 2:
The invention extracts the essential sensing function from native receptor systems by creating an artificial biosensor that uses only the ligand-binding domain for extracellular recognition, while the intracellular signaling function is provided by a separate, engineered functional domain. This extraction eliminates dependence on native signaling machinery and its associated interference.
2Adaptability or versatility
If reporter systems are transferred between cell types, then the system can be applied to different contexts, but native regulatory mechanisms interfere and complicate interpretation
Solution Approach 1:
The biosensor employs local quality by making the functional domain specific to a particular transcription factor and promoter pair. This localization of function ensures that the biosensor signal is directly interpretable in terms of ligand binding events, independent of the host cell's native regulatory landscape, thereby maintaining signal fidelity across different cell types.
3Reliability
If native receptors and signaling proteins are required, then the system can monitor native signaling, but the components must be present in the cell or exogenously expressed at high levels
Solution Approach 1:
The invention merges the ligand-binding function (from the extracellular domain) with the signaling function (from the intracellular functional domain) into a single chimeric receptor protein. This merging eliminates the need for separate native receptors and signaling proteins, simplifying system setup while maintaining reliable signal detection.
4Reliability
If TANGO biosensors are designed using native receptors and signaling proteins, then the system can monitor specific interactions, but identifying suitable components is time-consuming and limits detection of novel ligands
Solution Approach 1:
The biosensor system achieves universality by using a standardized intracellular functional domain (such as a transcription factor fused to a reporter gene) that can work with any ligand-binding domain. This allows rapid development of biosensors for novel ligands by simply swapping the extracellular ligand-binding domain, while the intracellular signaling module remains constant, dramatically reducing development time.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
MESA biosensors are highly customizable and efficient, capable of detecting ligands for which no natural receptor exists, with reduced background signaling and improved signal-to-noise ratio, allowing for robust and orthogonal sensing of extracellular cues.
Implementation Method 1
a protease domain, a protease cleavage site
Data Source
AI summary
The present invention provides modular extracellular sensors, nucleic acids encoding such sensors, and cells expressing such sensors, and methods of employing such sensors and cells for detecting extracellular ligands. In certain embodiments, the extracellular sensors comprise a ligand binding domain, a transmembrane domain, a protease domain, a protease cleavage site, and a transcription factor. In other embodiments, a pair of extracellular receptors is provided where both receptors contain a ligand binding domain and transmembrane domain, and one receptor contains a protease cleavage site and a transcription factor and the other receptor contains a protease domain.