mRNA Sequences with miRNA Binding Sites for Cell-Type Specific Expression
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Solution Overview
Problem
Current methods for delivering polynucleotide sequences, such as mRNA, to specific organs and tissues face challenges in achieving effective targeting and differential expression in various cell types, leading to limitations in therapeutic efficacy and increased side effects.
Innovation Solution
An isolated mRNA sequence is designed with coding sequences for polypeptides, including untranslated regions (UTRs) and multiple micro-RNA (miRNA) binding sites, allowing for differential expression in different cell types within target organs, combined with targeted delivery particles to enhance specificity and efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If non-viral methods are used to deliver polynucleotides to cells, then safety and reduced toxicity are improved, but transfection rates and targeting efficiency deteriorate
Solution Approach 1:
The patent modifies the mRNA structure by incorporating specific UTR sequences and miRNA binding sites, changing the biochemical parameters of the polynucleotide to enhance cell-type specific expression and transfection efficiency while maintaining safety
Solution Approach 2:
The patent uses miRNA binding sites as intermediaries that mediate between the delivered mRNA and the target cell's gene regulation machinery, enabling selective expression in desired cell types while avoiding toxicity in others
2Productivity
If viral vectors are used to deliver coding polynucleotides, then transfection rates and targeting ability are improved, but risks of toxicity and inflammation worsen
Solution Approach 1:
The patent employs transient mRNA expression instead of persistent viral integration, using short-lived mRNA molecules that provide temporary therapeutic effect without the long-term safety risks of viral vectors
Solution Approach 2:
The patent engineers specific mRNA structural parameters including UTR sequences and miRNA binding sites to achieve viral-level transfection efficiency and cell-type specificity without invoking the immune response associated with viral vectors
3Adaptability or versatility
If polynucleotides are delivered to achieve broad expression, then therapeutic coverage is improved, but side effects and off-target effects worsen
Solution Approach 1:
The patent confers cell-type specific expression by incorporating miRNA binding sites that create local regulatory differences in the mRNA, allowing the same polynucleotide to be expressed in some cell types while silenced in others, thereby achieving targeted therapy with reduced side effects
4Manufacturing precision
If differential expression in different cell types is achieved through targeted delivery, then therapeutic precision is improved, but delivery system complexity worsens
Solution Approach 1:
The patent extracts the targeting function from the delivery vehicle and embeds it directly into the mRNA molecule itself through incorporated miRNA binding sites, simplifying the overall delivery system while maintaining high targeting precision
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 enables precise and controlled expression of therapeutic agents in specific cell types, improving treatment outcomes while minimizing side effects by leveraging miRNA-mediated modulation for tissue-specific translation.
Implementation Method 1
The miRNA binding site sequences allow for differential expression of the coding sequence in at least a first and a second cell type within the target organ or organs
Data Source
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AI summary
Provided is an isolated mRNA sequence for expression of one or more cytokines within one or more target organs, the sequence comprising at least one coding sequence which codes for one or more cytokines (or their ligands) involved in immune response and inflammation selected from one or more of: TNFα, TNFβ, IFNα, IFNβ, IFNgamma, IL1, IL2, IL3, IL4, IL5, IL6, IL7, IL8, IL9, IL10, IL11, IL12, CCL2, CCL3, CCL4, CCL5, CXCL 9, and CXCL10. The sequence also comprises at least a first untranslated region (UTR) sequence, and at least three different micro-RNA (miRNA) binding site sequences. Each of the miRNA binding site sequences is located within, immediately 5' to or immediately 3' to, the first UTR sequence; and the miRNA binding site sequences allow for differential expression of the coding sequence in at least a first and a second cell type within the target organ or organs. Also provided are pharmaceutical compositions comprising the mRNA sequence, and methods for using the sequence or composition, particularly in treatment of disease, such as cancer of the liver, brain, lung, breast, pancreas, colon and kidney.