Polyamine-Responsive Splicing Elements for Fine-Tuned Gene Expression
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Solution Overview
Problem
There is a need for a versatile platform system to regulate gene expression in eukaryotic cells, particularly in plants, algae, fungi, and mammals, to control the expression of heterologous and endogenous genes in a cost-effective and reliable manner, including the fine-tuning of desired product production and prevention of undesired gene expression in transgenic organisms.
Innovation Solution
The system employs an expression control element (ECE) containing a polyamine or polyamine analog-responsive nucleic acid sequence flanked by splice sites, which mediates alternative splicing in response to polyamine or analog levels, allowing for precise regulation of RNA and protein expression through alternative splicing mechanisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If alternative splicing is used to regulate gene expression, then gene expression control is improved, but system complexity increases
Solution Approach 1:
The patent introduces an alternative splicing regulatory element (ASRE) as an intermediary component that mediates between the promoter and coding sequence. This ASRE element contains specific splice sites and regulatory sequences that control alternative splicing events, providing a modular and manageable mechanism for gene expression regulation without requiring complex overall system design
Solution Approach 2:
The gene expression system is divided into distinct functional segments: a promoter region, an alternative splicing regulatory element (ASRE) containing splice sites and regulatory sequences, and a coding sequence. This segmentation allows independent optimization and control of each component, simplifying the overall system while maintaining reliable gene expression control
2Manufacturing precision
If polyamine-responsive elements are used for gene regulation, then regulation precision is improved, but manufacturing complexity increases
Solution Approach 1:
The polyamine-responsive element functions autonomously by directly binding polyamine molecules to induce conformational changes that regulate splicing. This self-service mechanism eliminates the need for additional protein cofactors or complex regulatory machinery, achieving precise regulation while maintaining ease of manufacture through a single functional element
Solution Approach 2:
The system exploits changes in polyamine concentration as a regulatory parameter. By designing the ASRE element to respond to specific polyamine levels, the system achieves precise control over gene expression timing and intensity. The element undergoes conformational parameter changes when polyamines bind, switching between splicing states in response to physiological polyamine fluctuations
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 ECE effectively controls gene expression by enhancing or reducing the frequency of splicing, enabling fine-tuning of desired product production and preventing undesired gene expression, with applications in regulating resistance to herbicides, pesticides, and fungicides, and synchronizing gene expression with their application.
Implementation Method 1
The ECE comprises a polyamine or polyamine analog-responsive nucleic acid sequence flanked by splice sites or variants thereof
Data Source
AI summary
The present invention relates to an expression control element and expression system comprising same, particularly to an expression control element regulated by a polyamine or polyamine analogue, host cells and eukaryotic organisms comprising same and methods of use thereof.