Circular RNA Compositions for Non-Integrating Therapeutic Expression
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Solution Overview
Problem
Conventional DNA-based gene therapy methods risk integrating into the host genome, causing mutations, disrupting essential gene function, and triggering immune responses, while viral vectors are costly and difficult to deliver effectively.
Innovation Solution
The use of circular RNA polynucleotides, engineered with specific elements like IRES and aptamers, to facilitate expression of therapeutic proteins without integration into the genome, utilizing nanoparticle delivery systems for targeted cell entry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If DNA-based gene therapy is used to achieve long-lasting action, then the duration of therapeutic effect is improved, but the risk of genomic integration and harmful mutations increases
Solution Approach 1:
The patent extracts the therapeutic function from DNA and implements it using RNA molecules. The circular RNA molecules are designed to express therapeutic proteins without integrating into the host genome, thereby separating the beneficial long-lasting expression from the harmful integration risk. The circular structure enables sustained translation without requiring genomic incorporation.
Solution Approach 2:
The patent employs circular RNA molecules that are transient rather than permanent. These RNA molecules provide temporary but effective therapeutic expression, avoiding the need for permanent genomic integration. The circular structure protects them from degradation while allowing controlled, non-integrating expression of therapeutic proteins.
2Quantity of substance
If strong promoter sequences are included for effective gene expression, then the protein expression level is improved, but the risk of disrupting normal gene regulation increases
Solution Approach 1:
The patent extracts the expression function from DNA-based promoter systems and implements it using RNA-level mechanisms. The circular RNA molecules contain internal ribosome entry sites (IRES) and other RNA elements that directly control translation without requiring strong viral promoters, thereby achieving effective expression without disrupting host gene regulation.
Solution Approach 2:
The patent replaces the DNA-based promoter mechanism with an RNA-based translation control system. Instead of using strong promoters that bind transcription factors and regulate transcription, the circular RNA molecules use IRES elements and RNA secondary structures to directly control ribosome binding and translation initiation, achieving expression control at the translational level.
3Productivity
If viral vectors are used for delivery, then the efficiency of genetic material delivery is improved, but the cost and complexity of production increase
Solution Approach 1:
The patent uses circular RNA molecules as simplified copies of the therapeutic information carrier. Instead of using complex viral vectors that require extensive purification and quality control, the circular RNA can be produced through in vitro transcription from linear DNA templates, creating a simpler, more scalable system that maintains delivery effectiveness while reducing production complexity.
Solution Approach 2:
The patent replaces expensive, complex viral vectors with simpler circular RNA molecules that can be produced more economically. The circular RNA provides sufficient delivery efficiency for therapeutic purposes without requiring the complex viral capsid structures, enabling more cost-effective production while maintaining functional effectiveness.
4Measurement precision
If viral vectors are used for targeted delivery, then the precision of cell targeting is improved, but the risk of immune response increases
Solution Approach 1:
The patent extracts the targeting function from viral vectors and implements it using cell-penetrating peptides or ligand-conjugated systems attached to the circular RNA. This separates the delivery and targeting functions from the viral vector structure, achieving similar precision without triggering the strong immune responses associated with viral proteins.
Solution Approach 2:
The patent replaces the viral vector delivery system with a non-viral circular RNA system that uses alternative mechanisms for cellular entry and targeting. Instead of relying on viral envelope proteins and fusion mechanisms that trigger immune responses, the system uses chemically modified nucleotides, lipid conjugates, or peptide attachments to achieve targeted delivery without viral immunogenicity.
Data Source
AI summary
Circular RNA, along with related compositions and methods are described herein. In some embodiments, the inventive circular RNA comprises group I intron fragments, spacers, an IRES, duplex forming regions, and an expression sequence. In some embodiments, the expression sequence encodes an antigen. In some embodiments, circular RNA of the invention has improved expression, functional stability, immunogenicity, ease of manufacturing, and/or half-life when compared to linear RNA. In some embodiments, inventive methods and constructs result in improved circularization efficiency, splicing efficiency, and/or purity when compared to existing RNA circularization approaches.