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A Closed-Type Wireless Nanopore Electrode for Analyzing Single Nanoparticles
Published on: March 20, 2019
Nanopore label-free detection of single-nucleotide deletion in Baxα/BaxΔ2
Xiaohan Chen1, Liang Wang2, Golbarg M Roozbahani1
1Department of Chemistry, Illinois Institute of Technology, Chicago, IL, USA.
Abstract:
Baxα, a key tumor suppressor gene, will not be expressed correctly as a result of single nucleotide mutation in its microsatellite region; Instead, BaxΔ2, an isoform of Baxα, is often produced. In addition, lack of the exon 2 due to an alternative splicing, BaxΔ2 has the same sequence as Baxα except single base deletion from eight continuous guanines (G8) to G7. Most of the currently available methods for Bax∆2 detection are inefficient and time-consuming, and/or require the use of labels or dyes. In this work, we reported a label-free nanopore sensing strategy to differentiate between Baxα and BaxΔ2 with a DNA polymer as a molecular probe based on alternative spliced sequences. Two DNA molecules were designed to selectively detect Baxα and BaxΔ2, respectively. The method was rapid, accurate, and highly sensitive: picomolar concentrations of target nucleic acids could be detected in minutes. Our developed simple and fast nanopore-based detection strategy is not only useful for distinguishing between Baxα and Bax∆2, but also provides a useful tool for detection of other single-base mutations in genetic diagnosis.
Insights
This study introduces a novel, label-free nanopore sensing method to quickly and accurately detect BaxΔ2, an isoform of the tumor suppressor Baxα, caused by single-base mutations. This technique offers a sensitive tool for genetic diagnosis, distinguishing between Baxα and BaxΔ2 variants.
Area of Science:
- Molecular Biology
- Nanotechnology
- Genetics
Background:
- Baxα is a crucial tumor suppressor gene.
- Single nucleotide mutations in its microsatellite region can lead to the production of BaxΔ2, an alternatively spliced isoform.
- Current detection methods for BaxΔ2 are often inefficient, time-consuming, and require labels.
Purpose of the Study:
- To develop a rapid, label-free, and sensitive method for differentiating between Baxα and BaxΔ2.
- To utilize nanopore sensing with DNA probes for detecting single-base mutations.
Main Methods:
- A label-free nanopore sensing strategy was employed.
- DNA polymers were designed as molecular probes to selectively detect Baxα and BaxΔ2.
- The method relies on distinguishing sequence differences caused by alternative splicing.
Main Results:
- The developed strategy successfully differentiated between Baxα and BaxΔ2.
- The method demonstrated high sensitivity, detecting picomolar concentrations of target nucleic acids.
- Detection was achieved within minutes, showcasing rapidity.
Conclusions:
- A simple, fast, and sensitive nanopore-based detection strategy for Baxα and BaxΔ2 was established.
- This method provides a valuable tool for genetic diagnosis, particularly for single-base mutations.
- The approach avoids the need for labels or dyes, simplifying the detection process.
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