Functionalized Nanoneedle Array for Non-Destructive miRNA Detection
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Solution Overview
Problem
Current methods for determining microRNA in cells are time-consuming, costly, and often cause irreversible cell damage, limiting their suitability for high-throughput analysis and real-time monitoring of miRNA expression changes.
Innovation Solution
A method involving the delivery of probes into cells, followed by the insertion of functionalized nanoneedles to bind with target nucleic acids, and subsequent hybridization with complementary DNA sequences to produce a hybridized product, allowing for efficient and non-destructive determination of miRNA levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If qRT-PCR based miRNA kit is used to determine miRNA amount, then measurement precision is improved, but device complexity and time consumption increase
Solution Approach 1:
The method segments the detection process into distinct functional modules: probe delivery via nanoneedles, in-cell hybridization with target miRNA, and signal readout. This modular approach simplifies the overall system while maintaining measurement precision, eliminating the need for complex sample collection and testing steps required by qRT-PCR
Solution Approach 2:
The patent introduces probes as intermediary molecules that facilitate direct detection of miRNA within living cells. These probes serve as mediators between the detection system and the target miRNA, enabling precise measurement without the complex sample preparation steps required by conventional methods
2Measurement precision
If RNA sequencing is used for whole transcriptome profiling, then measurement precision is improved, but cost increases
Solution Approach 1:
The patent employs disposable probes that can be delivered into cells and used for detection. These single-use probes eliminate the need for expensive, maintainable sequencing platforms while achieving precise measurement of transcript levels through targeted hybridization rather than whole transcriptome sequencing
3Measurement precision
If nanoparticle introduction or probe insertion is used for single-cell analysis, then measurement precision is improved, but cell damage occurs
Solution Approach 1:
The patent replaces mechanical insertion methods with a chemistry-based approach where probes are delivered into cells through a non-mechanical process. This substitution eliminates the physical damage caused by nanoparticle or probe insertion while maintaining the ability to perform single-cell miRNA detection with high precision
4Measurement precision
If miRNA isolation from cell lysate is performed, then measurement precision is improved, but cell damage and loss of real-time monitoring capability occur
Solution Approach 1:
The patent performs preliminary hybridization of probes with target miRNA directly within living cells before any extraction or lysis steps. This preliminary action allows precise quantification of miRNA in its native cellular environment, enabling real-time monitoring without irreversible cell damage that occurs with traditional isolation methods
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 cost-effective, high-throughput analysis of miRNA levels without damaging cells, allowing for real-time monitoring and multiple sequence profiling, including during mouse embryonic stem cell differentiation.
Implementation Method 1
each of the one or more probes being capable of binding with corresponding target nucleic acid present in the cells to form a double-stranded sequence
Implementation Method 2
inserting an array of functionalized nanoneedles into the cells to bind with the double-stranded sequence
Implementation Method 3
hybridizing the bound double-stranded sequence with a first and second DNA sequence to produce a hybridized product, the first and second DNA sequence being at least partially complementary to each other
Data Source
AI summary
A method of determining one or more target nucleic acids in cells includes the steps of: delivering one or more probes into the cells, each of the one or more probes being capable of binding with corresponding target nucleic acid present in the cells to form a double-stranded sequence; inserting an array of functionalized nanoneedles into the cells to bind with the double-stranded sequence; and hybridizing the bound double-stranded sequence with a first and second DNA sequence to produce a hybridized product, the first and second DNA sequence being at least partially complementary to each other. A kit for determining a target nucleic acid in cells includes a first reagent comprising a probe for binding with the target nucleic acid to form a double-stranded sequence; and an array of functionalized nanoneedles comprising a protein for binding with the double-stranded sequence.