Yeast Colorimetric Reporter for Protein Aggregation Screening
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Solution Overview
Problem
Current cell-based systems for screening therapeutic agents for neurodegenerative diseases are limited by labor-intensive sample preparation and cost-consuming instruments, making it difficult to identify novel compounds that modulate protein aggregation effectively, especially for conditions like Huntington's disease where protein aggregation is a key factor.
Innovation Solution
A recombinant protein with a glutamine-rich domain and a color-alteration enzyme, such as ADE2, is used in yeast cells to create a color-based reporter system where the color change indicates protein aggregation status, allowing for easy detection of therapeutic agents that disrupt aggregation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional cell-based screening systems are used to identify therapeutic agents for neurodegenerative diseases, then the screening can detect protein aggregation, but the process becomes labor-intensive and requires expensive instruments
Solution Approach 1:
The patent employs a colorimetric reporter system where the ADE2 enzyme restores colony color when polyglutamine aggregates are disrupted. This visual color change provides a simple, instrument-free method to screen for compounds that modulate protein aggregation, eliminating the need for expensive equipment while maintaining detection capability
Solution Approach 2:
The invention uses a disposable yeast-based screening platform where each well contains yeast cells expressing the recombinant protein with polyglutamine tract. The yeast colonies serve as single-use, low-cost indicators that eliminate the need for expensive, reusable instrumentation, making high-throughput screening economically feasible
2Measurement precision
If traditional cell-based screening systems are used to identify therapeutic agents for neurodegenerative diseases, then the screening can detect protein aggregation, but the process becomes labor-intensive and requires cost-consuming instruments
Solution Approach 1:
The colorimetric output of the ADE2 enzyme provides a visually detectable signal that eliminates complex measurement procedures. Researchers can simply observe colony color to determine whether compounds disrupt protein aggregation, making the screening process accessible without specialized training or equipment
Solution Approach 2:
The yeast cells autonomously perform the detection function through the ADE2 enzyme's colorimetric activity. The system self-detects protein aggregation status without requiring external instruments or complex sample preparation, dramatically simplifying the screening workflow
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 system enables efficient screening of therapeutic agents by visually differentiating between aggregated and non-aggregated protein forms, facilitating the identification of compounds that reduce protein aggregation and potentially treat neurodegenerative diseases like Huntington's disease.
Implementation Method 1
A recombinant protein with a glutamine-rich domain and a color-alteration enzyme, such as ADE2, is used in yeast cells to create a color-based reporter system where the color change indicates protein aggregation status
Data Source
AI summary
This invention provides recombinant protein comprising formula as follows:A-Bwherein A is polypeptide containing glutamine-rich domain comprising 8-200 glutamine residues and B is color-alteration enzyme modulated by the domain. This invention also provides an expressing vector and a cell produced therefrom. This invention further provides a method and a kit for screening a therapeutic agent for a neurodegenerative disease.