Circular RNA Enrichment via Denaturing Conditions
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Solution Overview
Problem
Current methods for separating and purifying circular polyribonucleotides (circRNA) from mixed populations are inefficient, particularly in scaling up processes and achieving high purity, often relying on gel electrophoresis and not effectively addressing impurities.
Innovation Solution
The method involves exposing a sample containing circRNA and linear polyribonucleotides to denaturing conditions such as thermal denaturation, extreme pH, or chemical treatments to selectively enrich circRNA, avoiding the use of gel electrophoresis and facilitating large-scale purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If gel electrophoresis is used for circRNA separation, then separation capability is provided, but device complexity and process scalability are worsened
Solution Approach 1:
The patent replaces gel electrophoresis (a mechanical/electrical separation system) with a chemical denaturation approach. By treating RNA with denaturing agents under controlled conditions, circRNA is selectively enriched without requiring complex electrophoretic equipment, thereby simplifying the system while maintaining separation effectiveness
Solution Approach 2:
The patent utilizes changes in chemical parameters (pH, temperature, denaturing agent concentration) to achieve separation. By adjusting these parameters during the denaturation process, circRNA is selectively stabilized while linear RNA is degraded, providing an effective separation mechanism that avoids complex device requirements
2Manufacturing precision
If conventional purification methods are used, then circRNA enrichment is achieved, but productivity and scalability are worsened
Solution Approach 1:
The patent employs a self-service approach where the denaturing conditions themselves perform the separation function. The chemical and physical conditions automatically differentiate between circRNA and linear RNA based on their structural properties, eliminating the need for complex multi-step purification protocols and enabling scalable production
Solution Approach 2:
The patent exploits phase transition-like behavior in the denaturation process, where circRNA maintains its structured state while linear RNA transitions to a degraded state. This phase-like differentiation allows for efficient separation at scale, improving both productivity and purity simultaneously
3Manufacturing precision
If denaturing conditions are applied to enrich circRNA, then circRNA purity and yield are improved, but loss of substance may occur
Solution Approach 1:
The patent applies local quality differentiation by creating specific denaturing conditions that selectively affect linear RNA while preserving circRNA. The denaturing agents are applied in a controlled manner that targets the vulnerable linear structures while leaving the resilient circular circRNA intact, thereby achieving purity without excessive loss
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
This approach effectively increases the purity and yield of circRNA, allowing for the production of large quantities while reducing impurities, and is scalable for high-throughput applications.
Implementation Method 1
separation of circRNA a under denaturing conditions (e.g., thermal denaturation, pH, or chemical treatment)
Data Source
AI summary
The present disclosure is directed to methods for the enrichment of circular polyribonucleotides (circRNA), e.g., from population of polyribonucleotides containing circRNA and linear polyribonucleotides (linRNA), where the enrichment is performed under denaturing conditions. Also disclosed are compositions including a population of polyribonucleotides containing circRNA and linRNA in a solution under denaturing conditions. Further within the scope of the present disclosure are compositions containing an enriched population of circRNA, such as a composition that was produced by exposing the composition to one or more denaturing conditions.