Modified U7 snRNA for TDP-43 Cryptic Exon Splicing Repression
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Solution Overview
Problem
Loss of nuclear TDP-43 leads to the inclusion of cryptic exons in genes like STMN2 and UNC13A, resulting in functional protein depletion and aberrant RNA processing, contributing to neurodegenerative diseases such as ALS and FTD, with no effective therapeutic approaches to address this issue.
Innovation Solution
A modified U7 snRNA construct with antisense sequences complementary to TDP-43 regulated cryptic exons and flanking regions, blocking splicing machinery to prevent inclusion of cryptic exons, thereby restoring functional protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TDP-43 is depleted in cells, then cryptic exons are included in mature RNA, but functional protein expression is reduced
Solution Approach 1:
The patent introduces a modified U7 snRNA construct as an intermediary molecule that binds to cryptic exon sequences and recruits splicing repressors, mediating the suppression of cryptic exon inclusion in the absence of TDP-43. This intermediary mechanism restores proper splicing fidelity without requiring TDP-43 presence.
Solution Approach 2:
The invention changes the splicing parameters by introducing exogenous snRNA constructs with altered sequence specificity that target cryptic exons differently from endogenous splicing machinery. This parameter change enables selective suppression of cryptic exon inclusion while preserving normal exon splicing.
2Adaptability or versatility
If conventional therapeutic approaches are used, then general disease treatment is attempted, but no effective therapy exists for TDP-43 related neurodegenerative diseases
Solution Approach 1:
The modified U7 snRNA construct serves multiple functions: it acts as a guide RNA for sequence-specific binding, recruits splicing repressor proteins, and modulates splicing outcomes. This multi-functionality enables a single therapeutic agent to address multiple aspects of TDP-43-related splicing defects across different neurodegenerative diseases.
Solution Approach 2:
The snRNA construct utilizes the cell's own splicing machinery and endogenous splicing repressors to achieve therapeutic effects, rather than requiring external delivery of complex protein therapeutics. The system self-assembles and functions within the cellular environment using existing molecular pathways.
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 modified U7 snRNA construct effectively represses cryptic exon splicing, restoring normal protein production even in the absence of TDP-43, offering therapeutic potential for diseases associated with TDP-43 dysfunction.
Implementation Method 1
at least one antisense sequence having between 16 to 30 nucleotides which are at least 90% complementary to a splicing element of a TDP-43 regulated cryptic exon sequence
Data Source
AI summary
A modified U7 snRNA construct comprising at least one antisense sequence having between 16 to 30 nucleotides which are at least 90% complementary to a splicing element of a TDP-43 regulated cryptic exon sequence or flanking regions thereof and wherein the U7 snRNA construct is capable of modulating splicing of the TDP-43 regulated cryptic exon in a cell. The splicing element may be selected from a splice site, a TDP-43 binding region, or an exonic splice enhancer. Vectors and pharmaceutical compositions comprising the construct are also described, as well as constructs for use in the treatment of diseases associated with TDP-43 dysfunction. Example TDP-43 regulated cryptic exon sequences include TDP-43 regulated cryptic exons in UNC13A, STMN2 and INSR genes.