Activatable Promoters With Photocleavable Repression Control
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Solution Overview
Problem
Existing inducible transcription and translation systems lack stringent regulation between activated and repressed states, and are difficult to encapsulate in synthetic cells, limiting their application in synthetic biological systems.
Innovation Solution
Development of activatable promoters with photocleavable moieties that uncouple transcription inhibition moieties upon exposure to electromagnetic radiation, allowing for precise control of transcription and translation processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing inducible transcription systems are used, then transcription control is achieved, but regulation between activated and repressed states is not stringent
Solution Approach 1:
The patent extracts the transcription inhibition function into a separate, removable component (inhibitor molecule) that can be precisely controlled. By taking out the inhibition mechanism as a distinct element that can be added or removed, the system achieves stringent regulation - the inhibitor can be completely removed to activate transcription or precisely controlled to repress it, solving the problem of non-stringent regulation in existing systems.
Solution Approach 2:
The patent implements dynamic control by making the transcription inhibition state changeable through light irradiation. The inhibitor molecule's binding and unbinding states can be dynamically switched using photocleavable linkers, allowing the system to transition between repressed and activated states with precise temporal control, thereby improving both regulation stringency and control precision.
2Adaptability or versatility
If light activated transcription systems are used, then flexibility and control are improved, but the systems cannot readily be encapsulated inside certain types of synthetic cells
Solution Approach 1:
The patent uses an intermediary approach by incorporating the light-sensitive inhibitor complex within a protective capsule or synthetic cell structure. The photocleavable linker acts as an intermediary that allows light energy to penetrate through the capsule boundary to trigger inhibitor release inside the synthetic cell, thus maintaining flexibility and control while enabling encapsulation in synthetic cell systems.
3Reliability
If transcription inhibition moieties are coupled to the promoter, then repression is achieved, but uncoupling requires specific electromagnetic radiation
Solution Approach 1:
The patent applies parameter changes by using photocleavable linkers with specific wavelength sensitivities. By selecting linkers that respond to specific regions of the electromagnetic spectrum (e.g., UV, visible light), the system achieves reliable repression during normal conditions while enabling controlled uncoupling through targeted light irradiation. This parameter-based control allows differentiation between repression state and activation state without requiring complex additional mechanisms.
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
Provides a system with stringent regulation and encapsulation capabilities, enabling controlled expression of biological molecules in synthetic biological systems.
Implementation Method 1
The one or more cleavable moieties are preferably photocleavable moieties. The one or more photocleavable moieties may be a moiety that may be cleaved upon contact with electromagnetic radiation
Data Source
AI summary
The present invention relates to activatable promoters, as well as expression systems, vectors, delivery systems, reaction vessels, aqueous objects and compositions comprising said activatable promoters. The invention also provides compositions comprising assemblies and networks of aqueous objects comprising said activatable promoters. The invention additionally provides methods of expressing a transcript and methods of expressing a polypeptide using the activatable promoters of the invention. The invention further provides phospholipid mixtures, as well as aqueous objects and compositions comprising said phospholipid mixtures