Sensor Protein Expression Cassettes for Stable Chemical Response
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Solution Overview
Problem
Existing odor sensors that rely on lipid bilayer membranes with sensor proteins are inefficient to produce, and there is a need for improved production efficiency and stability of chemical response activity regardless of the type of sensor protein.
Innovation Solution
A polynucleotide containing an expression cassette with a coding sequence for a sensor protein and another cassette with coding sequences for additional proteins, such as chromogenic or luminescent proteins and drug-resistant genes, optimized for stable expression and activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiple protein-coding sequences are incorporated into a single expression cassette to improve efficiency, then the number of vectors and delivery steps is reduced, but the position of the sensor protein-coding sequence affects chemical response activity and some sensor proteins may not exhibit activity
Solution Approach 1:
The patent divides the expression system into two separate expression cassettes: Expression Cassette 1 contains only the sensor protein-coding sequence, while Expression Cassette 2 contains at least two other protein-coding sequences. This segmentation ensures that the sensor protein is expressed independently without being affected by positional effects relative to other sequences, thereby maintaining reliable chemical response activity while still achieving efficient co-expression of multiple proteins through a single vector.
2Device complexity
If multiple protein-coding sequences are placed in a single expression cassette to reduce vector size, then vector delivery efficiency is enhanced, but the sensor protein's chemical response activity becomes position-dependent
Solution Approach 1:
The patent segments the expression cassette into two independent units within a single vector: Expression Cassette 1 dedicated to sensor protein expression and Expression Cassette 2 for other proteins. This segmentation simplifies the structural design by using standard promoter-terminator units while ensuring reliable sensor protein activity through independent expression control.
Solution Approach 2:
The patent extracts the sensor protein-coding sequence from the group of multiple protein-coding sequences and places it in a separate expression cassette. This extraction eliminates the positional effects that would otherwise occur when the sensor protein sequence is embedded among other sequences, thereby ensuring consistent chemical response activity regardless of the presence or position of other genes.
3Productivity
If sensor cells are produced with multiple protein-coding sequences in a single expression cassette, then production efficiency improves, but stability of chemical response activity varies depending on sensor protein type and position
Solution Approach 1:
The patent implements segmentation by creating two separate expression cassettes within the vector: Expression Cassette 1 for sensor protein and Expression Cassette 2 for other proteins. This segmentation strategy enables stable and reliable production of sensor cells with consistent chemical response activity across different sensor protein types, while maintaining high production efficiency through single-vector delivery.
Data Source
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AI summary
A polynucleotide is provided that enables chemical response activity with enhanced stability regardless of the type of sensor protein while containing coding sequences for multiple proteins, including a sensor protein. The polynucleotide contains an expression cassette 1, which contains only a coding sequence of a sensor protein as a protein-coding sequence, and an expression cassette 2, which contains at least two coding sequences for proteins.