Empowering protein single-molecule sequencing: nanopore technology toward sensing gene sequences

Area of Science:

• Biophysics
• Molecular Biology
• Nanotechnology

Background:

• Investigating proteins at the single-molecule level is crucial for understanding their structure-function relationship.
• Traditional methods provide average effects, limiting insights into individual molecular behavior.
• Nanopore sensing offers a unique approach to characterizing single molecules.

Purpose of the Study:

• To review the sensing mechanisms of nanopore sequencing technology for various biological applications.
• To highlight the advantages and applications of nanopore sequencing in biological and solid-state systems.
• To discuss current challenges and future directions in nanopore-based single-molecule analysis.

Main Methods:

• Review of sensing mechanisms in DNA damage, DNA methylation, RNA sequencing, and protein analysis (modifications, unfolding).
• Exploration of both biological and solid-state nanopore systems.
• Analysis of current changes (intensity, frequency, period) for molecular characterization.

Main Results:

• Nanopore sequencing provides single-molecule resolution, unlike ensemble techniques.
• Key advantages include fast readout, high accuracy, low cost, and portability.
• Applications span DNA-protein interactions, protein conformation, DNA sequencing, microbial assays, and clinical diagnostics.

Conclusions:

• Nanopore sequencing offers novel insights into complex biological processes and real-time molecular change monitoring.
• Its versatility extends to clinical diagnostics, environmental monitoring, food safety, and forensic analysis.
• Addressing challenges in reagents and device design will further enhance nanopore sensing for protein analysis and structure prediction.