miRNA Binding Sites for Cell-Type Specific mRNA Expression
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current RNA-based therapeutic delivery methods face challenges in maintaining on-target activities and tumor- or cell-specificity, leading to off-target side effects due to accumulation in unintended organs and tissues, such as the liver and kidney.
Innovation Solution
Incorporating specific miRNA binding sites, like miRNA-122, 148a, 101, and 192, into the 5' UTR or coding sequence of mRNA constructs to reduce expression in hepatocytes and other non-target cells, thereby enhancing cell-type specific expression and minimizing off-target effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If RNA-based therapeutics are delivered using lipid-based carrier systems, then intracellular delivery and effectiveness are improved, but accumulation in unintended organs and tissues (liver and kidney) occurs, leading to off-target side effects
Solution Approach 1:
The patent applies local quality by incorporating tissue-specific miRNA binding sites (e.g., miR-122 for liver, miR-148a for kidney) into the 3' UTR of mRNA constructs. This allows the RNA therapeutic to have different binding affinities in different tissues, enabling selective silencing of off-target expression in liver and kidney while maintaining on-target activity in the intended tissue.
Solution Approach 2:
The patent uses miRNA binding sites as intermediary elements that mediate between the mRNA construct and the cellular machinery. These binding sites interact with tissue-specific miRNAs to regulate translation, serving as a molecular bridge that enables selective gene expression control without altering the core therapeutic function.
2Reliability
If miRNA binding sites are incorporated into the 3' UTR to reduce off-target expression, then cell-type specificity is improved, but the complexity of the nucleic acid sequence increases
Solution Approach 1:
The patent segments the regulation function by placing miRNA binding sites specifically in the 3' UTR region, separating the therapeutic coding sequence from the regulatory elements. This segmentation allows the coding region to remain simple and focused on the therapeutic protein, while the 3' UTR contains the tissue-specific miRNA binding sites that provide selective expression control.
Solution Approach 2:
The patent employs partial action by incorporating only the necessary miRNA binding sites required for achieving the desired tissue specificity. Rather than adding multiple regulatory elements, the invention uses strategically selected binding sites in the 3' UTR that provide sufficient tissue-specific regulation with minimal sequence complexity.
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 incorporation of miRNA binding sites allows for targeted expression within specific organs and tissues, reducing unwanted expression in liver and immune cells, thereby enhancing the therapeutic efficacy and safety of RNA-based therapies.
Implementation Method 1
They function via Watson-Crick base-pairing with complementary sequences within the 3′ untranslated regions (3′ UTR) of target mRNA molecules
Data Source
AI summary
The present invention provides a nucleic acid sequence comprising at least one miRNA binding site sequence containing at least one miRNA binding site. Those miRNA binding site sequences are located within and/or immediately 3 or 5′ of the 5′ UTR of a gene to reduce the off-target side effects and allow a cell type specific expression from the nucleic acid sequence within the target organ or organs. The invention further provides pharmaceutical compositions, as well as a method of promoting cell-type specific expression, comprising the nucleic acid sequence according to the invention for use in therapy.