Recombinant miRNA Plasmid Composition for Checkpoint mRNA Silencing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Bioactive molecules such as immune checkpoint molecules are often overexpressed or misexpressed, leading to homeostasis disruption and associated diseases.
Innovation Solution
Compositions comprising recombinant plasmids that encode for microRNA sequences targeting the mRNA of immune checkpoint molecules like PD-1, PD-L1, PD-L2, CTLA4, or IDO1, increasing endogenous production of miRNA to degrade or inactivate these molecules, thereby reducing their bioavailability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If immune checkpoint molecules are overexpressed or misexpressed, then homeostatic control is disrupted and disease occurs, but increasing miRNA production to degrade these molecules may affect normal immune regulation
Solution Approach 1:
The patent applies local quality by designing miRNA molecules with specific sequence complementarity to target checkpoint molecules. Each miRNA is engineered to recognize and bind to specific mRNA sequences of overexpressed checkpoint molecules (PD-1, PD-L1, CTLA-4, etc.), allowing selective degradation of harmful overexpressed proteins while leaving normal regulatory mechanisms intact. This localized targeting ensures that only the pathological overexpression is suppressed, preserving normal immune homeostasis.
Solution Approach 2:
The patent utilizes parameter changes by modifying the expression levels of miRNAs through recombinant plasmid delivery. The system changes the concentration and activity parameters of specific miRNAs within target cells, increasing miRNA production to therapeutic levels that can effectively degrade overexpressed checkpoint molecules. This parameter modulation allows precise control over the degradation process, suppressing pathological overexpression while maintaining safe operational thresholds for normal immune function.
2Reliability
If recombinant plasmids are used to increase endogenous miRNA production, then expression of target biomolecules is downregulated, but the complexity of the composition increases
Solution Approach 1:
The patent applies preliminary action by incorporating pre-designed recombinant plasmids that contain the necessary genetic elements for miRNA expression before administration to the subject. The plasmids are engineered in advance with promoter sequences, miRNA coding regions, and regulatory elements optimized for efficient transcription and translation. This preliminary preparation ensures that upon delivery, the plasmids can immediately begin producing therapeutic miRNAs without requiring complex in vivo assembly or processing steps, thereby simplifying the overall composition while achieving reliable downregulation.
3Productivity
If miRNA sequences are designed to be complementary to target mRNA, then translation of target biomolecule is inhibited, but specificity of targeting must be maintained to avoid off-target effects
Solution Approach 1:
The patent applies segmentation by dividing the target mRNA binding interface into multiple discrete miRNA sequences, each with specific complementarity to different regions of the target mRNA. Instead of using a single long complementary sequence, the system employs multiple shorter miRNA molecules (e.g., miR-34a, miR-124, miR-200 family members) that collectively target different segments of overexpressed checkpoint molecule mRNAs. This segmentation approach increases the probability of achieving high-specificity binding while reducing the risk of off-target effects, as each individual miRNA has a more restricted binding window.
Solution Approach 2:
The patent incorporates feedback mechanisms by designing miRNA sequences and delivery systems that respond to the actual expression levels of target checkpoint molecules. The recombinant plasmids are constructed with regulatory elements that can sense and respond to cellular conditions, adjusting miRNA production rates based on the presence and abundance of target mRNA. This feedback control ensures that miRNA degradation activity is proportional to the pathological overexpression level, maintaining high productivity when needed while automatically reducing activity when target levels are normal, thereby preserving manufacturing precision and avoiding off-target effects.
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 compositions effectively downregulate the expression of immune checkpoint molecules, potentially treating or preventing diseases related to their overexpression or misexpression by enhancing miRNA production within cells.
Implementation Method 1
The sequences of miRNA may be complimentary to a sequence of target messenger RNA (mRNA) that encodes for translation of a target biomolecule and the miRNA can cause the target mRNA to be degraded or inactivated
Data Source
AI summary
Embodiments of the present disclosure relate to a composition that comprises a recombinant plasmid (RP) with a sequence of nucleic acids. The sequence comprise a start region, an end region and an insert positioned between the start region and the end region. The insert encodes for a sequence of micro interfering ribonucleic acid (miRNA) that may be complimentary to a sequence of target messenger RNA (mRNA) that encodes for translation of a target biomolecule. The miRNA can cause the target mRNA to be degraded or inactivated, thereby causing a decrease in bioavailability of the target biomolecule because it is degraded or inactivated by the miRNA, thereby decreasing the bioavailability of the target biomolecule. In some embodiments of the present disclosure, the target biomolecule is an immune checkpoint protein.