Signal Activatable Molecular Constructs for Controlled Biomolecule Delivery
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Solution Overview
Problem
Current methods for controlled delivery of biomolecules to specific environments, such as cells or tissues, face challenges in achieving precise and controlled release, particularly in medical and biological applications.
Innovation Solution
The development of signal activatable constructs comprising a sensor domain with a protection segment, activation segment, displacement segment, and toehold segment, which upon binding of a signal polynucleotide, undergoes a conformational change to release a targeting domain via RNAse H cleavage, allowing for controlled and cell-type specific molecular delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional delivery methods are used, then delivery of biomolecules can be achieved, but controlled release to specific environments remains challenging
Solution Approach 1:
The delivery construct is segmented into distinct functional domains: a targeting domain for cell-specific delivery, a sensor domain with multiple segments (protection, activation, displacement, toehold) for signal-responsive control, and a cargo domain for the delivered biomolecule. This segmentation enables independent optimization of delivery precision and control mechanisms without overwhelming complexity.
Solution Approach 2:
The sensor domain undergoes dynamic conformational changes in response to cellular signals. The displacement segment and toehold segment interact to trigger a transition from an inactive state (where the targeting domain is bound) to an active state (where the targeting domain is released), enabling precise temporal control of biomolecule delivery based on real-time signal detection.
2Adaptability or versatility
If signal-responsive control is implemented, then cell-type specific modulation is enabled, but the molecular complex structure becomes more complex
Solution Approach 1:
The sensor domain is designed as a multi-functional module that integrates signal detection (toehold segment), conformational switching (displacement segment), and targeted release (activation segment with RNAse H site) into a single protein-free domain. This universal design allows the same sensor domain structure to respond to various cellular signals and control release of different cargo molecules, reducing overall system complexity while enhancing adaptability.
Solution Approach 2:
The displacement segment acts as an intermediary element that translates external cellular signals into internal conformational changes. When a signal binds to the toehold segment, the displacement segment mediates the structural transition that ultimately triggers RNAse H-mediated cleavage and release of the targeting domain, providing a controlled intermediate step between signal detection and cargo release.
3Stability of the object's composition
If the targeting domain is always bound to the protection segment, then stability is maintained, but controlled release cannot be achieved
Solution Approach 1:
The construct is pre-assembled with the targeting domain covalently bound to the protection segment in a stable inactive state, ensuring proper folding and stability during circulation or storage. The sensor domain components (displacement and toehold segments) are pre-positioned to enable rapid signal-responsive release, combining preliminary stabilization with preparedness for controlled activation when needed.
Solution Approach 2:
The system utilizes changes in molecular binding parameters (affinity, conformational energy states) to switch between stable bound and released states. The displacement segment and toehold segment are configured so that signal binding alters the thermodynamic parameters of the complex, triggering a phase transition from a stable bound state to a released state, thereby achieving controlled release while maintaining stability when required.
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 precise and efficient delivery of biomolecules by switching between inactive and active conformations, facilitating cell-type specific modulation and enhanced signal transduction, suitable for medical, research, and therapeutic applications like gene therapy and RNAi.
Implementation Method 1
the toehold segment is complementary to a signal polynucleotide, and the protection segment, displacement segment, and activation segment are configured so that upon complementary binding of the signal polynucleotide to the toehold segment
Implementation Method 2
the DNA portion of the activation segment complementarily binds the RNA portion of the protection segment to provide an RNAase H binding site presented for binding
Implementation Method 3
upon cleavage of the RNAase H binding site
Data Source
AI summary
Provided herein are signal activatable molecular constructs for enzyme-assisted delivery of molecules and related components, such as a sensor domain, compositions, methods and systems.