Artificial mRNA miRNA-Responsive Gene Expression Switch
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Solution Overview
Problem
Current mRNA-based systems lack an ON switch that can turn gene expression ON in response to specific miRNAs, making it difficult to target and kill cancer cells by activating apoptotic genes, and existing systems are complex due to the need for multiple components and foreign protein expression.
Innovation Solution
Development of artificial mRNA with a modified 3′-terminus structure, incorporating a miRNA target sequence and a translational repression sequence, allowing for ON switch functionality where protein expression is activated in response to specific miRNAs, enabling targeted gene expression in cells with high miRNA activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If a pseudo-ON switch is constructed using multiple OFF switches and foreign proteins, then gene expression can be turned ON in response to miRNA, but the system becomes complicated with multiple components and increased uncertainties
Solution Approach 1:
The invention extracts and utilizes the endogenous miRNA machinery already present in cells, eliminating the need for foreign protein expression systems. By designing mRNA molecules with specific miRNA target sequences and translational repression sequences, the system achieves miRNA-responsive gene expression using only cellular components, thereby reducing system complexity while maintaining adaptability.
Solution Approach 2:
The mRNA molecules are designed to self-regulate their translation through interaction with endogenous miRNAs. The translational repression sequence automatically responds to miRNA presence without requiring external control systems or foreign proteins, allowing the system to serve itself and reducing overall complexity.
2Adaptability or versatility
If existing OFF switch technology is used, then gene expression can be suppressed in response to miRNA, but it is impossible to turn gene expression ON to target and kill cancer cells
Solution Approach 1:
The invention inverts the logic of existing OFF switches by designing a system where miRNA binding activates rather than suppresses gene expression. By combining a miRNA target sequence with a translational repression sequence that is released upon miRNA binding, the system achieves ON-switch functionality, enabling activation of apoptotic genes in cancer cells that express specific miRNAs.
3Reliability
If multiple components are used to create an ON switch system, then miRNA-responsive expression is achieved, but the number of components increases and uncertainties increase
Solution Approach 1:
The invention merges the miRNA target sequence and translational repression sequence into a single mRNA molecule structure. This integration allows the mRNA to function as both the target for miRNA binding and the template for protein synthesis, eliminating the need for separate regulatory components and reducing overall system complexity while maintaining reliable miRNA-responsive expression.
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
Enables specific and controlled gene expression in cells with high miRNA activity, allowing for targeted cell killing and differentiation, and can be combined with existing OFF switches for versatile miRNA-directed gene control, suitable for medical applications.
Implementation Method 1
miRNA regulates decomposition of the corresponding mRNA, so that the miRNA controls gene expression
Data Source
AI summary
An mRNA forcibly expresses a protein gene in response to a miRNA, and a method for forcibly expressing the same, are provided. An artificial mRNA comprising a sequence encoding a protein gene, a miRNA target sequence linked to the 3′-terminal side of a Poly A sequence, and a translational repression sequence linked to the 3′-terminal side of the miRNA target sequence; and a method for expressing a protein gene in response to the expression of a miRNA, comprising a step of introducing the artificial mRNA into a cell.