U2OS Cell Expression of T2R Bitter Taste Receptors

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Solution Overview

Problem

Current methods for expressing T2R bitter taste receptors in HEK-293 cells result in a narrow signal window, limiting the ability to assess the interaction of test compounds across a broad range of activity levels, necessitating the development of alternative cell types for broader signal detection.

Innovation Solution

Expressing T2R bitter taste receptors in U2OS cells, which can include coupled G proteins such as gustducin or promiscuous G proteins like G15 or G16gust25, to provide a broader signal window for assessing test compound interactions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If T2R bitter taste receptors are expressed in HEK-293 cells, then the assay can detect bitter taste modulators, but the signal window remains narrow limiting assessment across broad activity levels

Engineering Contradiction:

Improvesignal window breadthVSAvoidrange of activity levels assessable

Core Design Contradiction:

Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The patent changes the cell type parameter from HEK-293 to U2OS cells, which fundamentally alters the signaling characteristics and expands the signal window. This parameter change enables assessment of test compounds across a broader range of activity levels while maintaining detection capability for bitter taste modulators.

Inventive Principle:

Principle #35Parameter changes

2Ease of manufacture

If T2R bitter taste receptors are expressed in HEK-293 cells, then the assay system is established, but the signal window is narrow reducing assay utility

Engineering Contradiction:

Improveassay establishmentVSAvoidsignal window

Core Design Contradiction:

Ease of manufactureVSMeasurement precision

Solution Approach 1:

The invention changes the cell line parameter from HEK-293 to U2OS cells, which provides a broader signal window while maintaining ease of assay establishment. The U2OS cells express T2R receptors with enhanced signaling properties that expand the measurable range without complicating the assay setup process.

Inventive Principle:

Principle #35Parameter changes

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

The use of U2OS cells with T2R bitter taste receptors and associated G proteins enhances the detection of bitter taste modulators and blockers, allowing for more accurate identification and reduction of bitterness in ingestible compositions.

Implementation Method 1

Taste receptors can be functionally expressed in cells, such as human embryonic kidney 293 (HEK-293) cells. Such taste receptors are coupled to one or more G-proteins. When a test molecule binds to the taste receptor, the coupled G-protein responds to the binding event by emitting a response, such as a change in the calcium cation concentration in the cell.

Methodology Applied

Scientific EffectG-protein coupled signaling:

Data Source

PatentUS20250102504A1Cells expressing bitter taste receptors and uses thereof

Publication Date: 2025.03.27 FIRMENICH INC

AI summary

The present disclosure generally relates to the expression of T2R taste receptors in eukaryotic cells, such as U2OS cells. The disclosure also relates to methods of expressing T2R taste receptors in such cells, and to methods of screening for substances that modulate T2R receptors, and which, therefore, may be useful as modulators of bitter taste.