miR183 Target Site Regulatory Sequences for Selective Gene Silencing
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Solution Overview
Problem
Current gene therapy approaches face challenges in ensuring precise expression of introduced genes only in specific cells and tissues to avoid deleterious effects, particularly in treating sensory impairments like blindness and deafness.
Innovation Solution
Utilization of vectors containing regulatory sequences with precursor miR183 target sites, which are complementary to miR183 family members, to selectively repress gene expression in cells where expression could be harmful, using nucleic acid sequences with at least 90% identity to SEQ ID NO: 1, SEQ ID NO: 21, or SEQ ID NO: 24.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If gene therapy approaches are used to treat sensory impairments, then gene expression can be introduced to compensate for defective genes, but the gene may be expressed in cells and tissues where it could induce deleterious effects
Solution Approach 1:
The patent uses miRNA target sites as intermediary elements that mediate between the introduced gene and the host cell's miRNA expression profile. These target sites act as molecular sensors that detect the presence of specific miRNAs (like miR183, miR96, miR182) and trigger selective gene silencing only in cells where these miRNAs are naturally expressed, thereby preventing deleterious effects while maintaining therapeutic benefits
Solution Approach 2:
The patent changes the regulatory parameters of gene expression by incorporating specific miRNA target sites into the gene construct. This modifies the expression profile from constitutive or non-specific expression to condition-specific expression based on miRNA presence, allowing the gene to be expressed only in appropriate cell types and tissues where the corresponding miRNAs are not present or present at low levels
2Adaptability or versatility
If regulatory sequences with miRNA target sites are used to control gene expression, then expression can be repressed in specific cells, but the complexity of the vector increases
Solution Approach 1:
The patent segments the regulatory control function into discrete miRNA target site modules that can be independently designed and inserted into the gene construct. Each target site corresponds to a specific miRNA, allowing modular assembly of specificity controls without requiring complete redesign of the entire vector system
Solution Approach 2:
The patent employs miRNA target sites that serve multiple functions: they act as silencing elements, provide cell-type specificity, and can be combined with different promoters and gene sequences. The same target site design principles apply across different gene therapy applications, making the approach universally applicable while maintaining relative simplicity
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 vectors effectively repress unwanted gene expression in sensory cells, providing a tool for targeted gene therapy by significantly reducing expression in cells expressing miR183, miR182, or miR96, enhancing specificity and safety.
Implementation Method 1
mature miRNAs achieve complete or incomplete base complementary pairing, which leads to mRNA cleavage and inhibition of mRNA translation into protein
Data Source
AI summary
An isolated nucleic acid sequence including at least two copies of a miRNA target site of the miR183 family having a sequence as set forth in SEQ ID NO: 1, SEQ ID NO: 21, or SEQ ID NO: 24, or a sequence having at least 90% identity with any one of SEQ ID NO: 1, SEQ ID NO: 21, or SEQ ID NO: 24. Also a vector including a regulatory element having at least one copy of this miRNA target site of the miR183 family. The isolated nucleic acid sequence and the vector may be particularly useful for controlling the expression of a gene of interest, for example when designing and developing gene therapies.