Closed-Loop Genetic Circuits for Stable Tunable Gene Expression
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Solution Overview
Problem
Current gene expression systems suffer from basal leakiness, lack of tunability, and lack of robustness, making them unsuitable for applications requiring precise and stable gene expression control, especially in biomanufacturing, biosensing, and ex-vivo therapies.
Innovation Solution
Engineered genetic circuits with closed-loop configurations, utilizing recombinant proteins and nucleic acid components that form feedback loops to regulate gene expression, including transcriptional activators and controllers, ensuring precise and stable expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If open-loop gene expression systems are used, then the system is simple to implement, but the expression level is vulnerable to perturbations and lacks robustness
Solution Approach 1:
The patent implements closed-loop genetic circuits where the output of one component feeds back to regulate earlier components. Specifically, a controller biomolecule is expressed in response to the output species, and this controller negatively regulates the activator or the output species itself, creating a feedback mechanism that stabilizes expression levels against environmental perturbations while maintaining system functionality.
2Ease of operation
If basal expression is allowed to occur, then the gene is always active, but leakiness occurs when expression should be silenced
Solution Approach 1:
The patent employs a controller biomolecule that is constitutively expressed and continuously acts to suppress basal leakiness. This controller negatively regulates the activator or output species before unwanted expression can occur, providing preemptive suppression of leaky expression while allowing full activation when the inducer is present.
3Quantity of substance
If the inducer concentration is increased to achieve higher expression, then the expression level increases, but the system loses tunability at specific levels
Solution Approach 1:
The patent creates a dynamic control system where the controller biomolecule concentration adjusts in response to the output species level. This dynamic feedback allows the system to achieve specific tunable expression levels by adjusting the controller's regulatory activity, enabling precise control over the range of expression levels rather than relying solely on fixed inducer concentrations.
4Reliability
If the expression system is made more robust to environmental fluctuations, then stability improves, but the system complexity increases
Solution Approach 1:
The patent implements a self-regulating system where the controller biomolecule automatically adjusts its regulatory activity in response to changes in output species concentration or environmental conditions. This self-service mechanism provides robustness against environmental fluctuations without requiring external control systems, as the circuit self-corrects to maintain stable expression levels.
Data Source
AI summary
The present invention is directed to engineered genetic circuits for controlling gene expression in cells, cells comprising said genetic circuits and uses of the cells and genetic circuits.