Nucleic Acid Probe for Splicing Protein Detection
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Solution Overview
Problem
Current diagnostic procedures for neurodegenerative disorders, such as ALS and Alzheimer's disease, are invasive, costly, and often detect protein misfolding late in disease progression due to low protein levels and reliance on clinical presentation, lacking efficient methods for monitoring splicing proteins' functional roles.
Innovation Solution
A nucleic acid probe with a pseudo intron and repeat nucleotide motifs for binding with splicing proteins, such as TDP-43 and FUS, is used in a homogeneous assay to detect functional splicing activity in biofluids, generating a signal upon pseudo intron excision and ligation of pseudo exons, allowing for quantitative analysis without lengthy wash steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current diagnostic procedures are used to detect splicing proteins, then detection can be performed, but the procedures are invasive, costly, and detect protein misfolding late in disease progression
Solution Approach 1:
The patent uses a nucleic acid probe as an intermediary substance that can be detected in biofluids to indirectly measure splicing protein activity. Instead of directly detecting the splicing proteins themselves (which requires invasive procedures), the probe serves as a mediator that translates protein function into detectable nucleic acid signals, enabling non-invasive monitoring through simple blood or urine collection
Solution Approach 2:
The invention replaces complex mechanical/invasive sampling procedures with a biochemical detection system. Instead of requiring tissue biopsies or invasive sampling to detect splicing proteins, the system uses in vitro splicing reactions with nucleic acid probes that can be analyzed from easily obtained biofluids, substituting mechanical intrusion with biochemical measurement
2Reliability
If current diagnostic procedures are used, then protein detection is possible, but it is costly and requires lengthy processing
Solution Approach 1:
The nucleic acid probe is pre-designed with built-in signal generation capabilities and recognition elements for splicing proteins. The probe undergoes preliminary preparation during synthesis, incorporating fluorophores, quenchers, or other detection moieties, so that when introduced to the sample, it requires minimal additional processing steps and can be detected directly, eliminating lengthy wash and handling procedures
Solution Approach 2:
The splicing reaction itself performs the detection function - the probe automatically undergoes splicing when the target protein is present, generating a detectable signal without requiring external intervention. The system is self-activating, where the biological activity being measured directly produces the detection signal, eliminating the need for separate amplification or processing steps
3Measurement precision
If invasive procedures are used for early detection, then sensitivity may be improved, but patient comfort and accessibility deteriorate
Solution Approach 1:
The nucleic acid probe acts as a sensitive intermediary that amplifies the detection signal for early disease stages. By measuring splicing activity through probe-based assays in biofluids, the system achieves high sensitivity comparable to invasive methods while using non-invasive sampling, as the probe selectively captures and amplifies signals from low levels of abnormal splicing activity
Solution Approach 2:
The invention changes the detection parameter from direct protein measurement (requiring invasive sampling) to nucleic acid-based splicing activity measurement (detectable in biofluids). This parameter transformation maintains sensitivity for early detection while improving accessibility, as nucleic acid signals from splicing reactions can be detected in easily obtained blood, urine, or other biofluids
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
Enables efficient monitoring of splicing proteins in accessible biofluids, providing a non-invasive, cost-effective method for early detection and quantification of functional splicing activity, potentially aiding in the assessment of disease progression and treatment efficacy.
Implementation Method 1
The probe comprises a repeat nucleotide motif as a recognition element for binding with a splicing protein
Implementation Method 2
the probe comprising a signal generating component for generation of a signal upon excision of the pseudo intron and ligation of the first pseudo exon and the second pseudo exon
Data Source
AI summary
The present invention relates to a nucleic acid probe for use in the detection of a functional splicing protein. The invention also relates to a method for the detection of a functional splicing protein, and a kit for the same. The invention further provides methods to quantify the presence of the level of functional protein in the sample.