Olfactory Receptor Expression for Sensitive Ligand Screening
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Solution Overview
Problem
Current methods for expressing olfactory receptors in host cells are limited in sensitivity, often requiring high ligand concentrations and are not compatible with high-throughput screening, particularly for poorly soluble and cytotoxic ligands, leading to incomplete ligand screening and false-positive results.
Innovation Solution
The use of modified olfactory receptor proteins with a specific C-terminal domain sequence motif, such as RN[KR][EDQ][VMIL][KR]xA[LIV][KRH][KR][LI][LIF][KRG][KR][KR], and optimized expression systems with accessory proteins like RTP1S and RTP2, enable uniform functional expression of a diverse repertoire of olfactory receptors, allowing sensitive detection at physiologically relevant concentrations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional expression systems are used for olfactory receptors, then expression can be achieved, but sensitivity is limited and high ligand concentrations are required
Solution Approach 1:
The patent modifies the C-terminal domain sequence of olfactory receptors to optimize their expression and detection properties. By changing the amino acid sequence parameters at the C-terminus, the system achieves enhanced sensitivity that allows detection at physiologically relevant ligand concentrations rather than requiring high concentrations
Solution Approach 2:
The patent introduces accessory proteins (RTP1S and RTP2) as intermediary components that facilitate proper folding, trafficking, and surface expression of olfactory receptors. These intermediary proteins enhance the functional expression of ORs, thereby improving detection sensitivity without requiring high ligand concentrations
2Productivity
If conventional screening methods are used, then some ligands can be detected, but high-throughput screening is not compatible and false positives occur
Solution Approach 1:
The modified C-terminal domain creates a more robust and uniform receptor expression system that reduces background noise and variability. This parameter change enables reliable high-throughput screening by maintaining detection accuracy even when processing large numbers of samples rapidly
Solution Approach 2:
The optimized expression system with accessory proteins creates a self-correcting system that naturally reduces false positives through improved receptor folding and trafficking. The system self-regulates to provide consistent, reliable signals that are compatible with automated high-throughput screening workflows
3Ease of manufacture
If standard olfactory receptor sequences are used, then expression is difficult to achieve, but uniform functional expression across diverse receptors is required
Solution Approach 1:
The patent applies a universal modified C-terminal domain sequence that can be used across diverse olfactory receptor variants. This universal modification works consistently across different OR classes and variants, enabling uniform functional expression while maintaining ease of manufacture through a standardized approach
Solution Approach 2:
Accessory proteins RTP1S and RTP2 act as intermediary chaperones that assist in the proper folding and trafficking of diverse olfactory receptor variants. These intermediary proteins compensate for sequence variations across different ORs, ensuring uniform functional expression across the entire repertoire of olfactory receptors
Data Source
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AI summary
Described herein are computer-implemented techniques for olfactory property prediction and olfactory receptor activation data prediction and use thereof in in the creation, modification, or improvement of flavours and fragrances.