Genetic regulatory networks implemented in various synthetic gene circuits in living cells for purpose of sensing, computing, and actuating in fields of diagnostics, monitoring etc. Untight control of transcriptional regulatory networks applied on promoters, which are the basic transcriptional regulatory component in synthetic circuits, often exhibit high basal level expression or leaky expression of target promoter. Most of the current solutions focus on other transcriptional and translational regulatory elements rather then on the sensing part of the system, which is the promoter itself. Furthermore, these methods affect the whole transcriptional unit in general, regardless the presence or absence of the input molecule, which result in simultaneous reduction of the minimum activity (basal level) together with the maximum activity of the promoter. The invention consists of a protocol that includes five steps which guarantees lowering the basal level expression and keeping high expression of a target promoter, resulting in an improvement in the fold change activation of promoter under test.
- Reduce the basal level of promoter without changing its maximal level.
- Modular systems
- Circuits can be implemented in whole-cell bacterial sensors.
Applications and Opportunities
- Environmental monitoring
- Metabolic engineering
- Therapeutics and Medical diagnosis
- Cell free systems
- Cellular immunotherapy