Advancing DNA and RNA Modification Detection via Nanopore Sequencing

DNA/RNA
modifications are crucial for biological processes. To
understand their regulatory mechanisms, precise mapping and quantification
of these modifications are essential. Although next-generation sequencing
can detect the location and stoichiometry of modifications, complex
treatments are required, hindering efficiency and long-range analysis.
In contrast, nanopore sequencing eliminates the need for additional
treatment and PCR amplification, thus preserving DNA/RNA modification
information. It enables modifications to be identified and quantified
directly at the single-molecule level. Therefore, nanopore sequencing
facilitates long-read, real-time modification detection, and has gained
widespread applications. This perspective introduces the principles
of nanopore sequencing, evaluates its strengths/weaknesses, and critically
examines its broader real-world applications. To expand the utility
of nanopore sequencing further, we discuss the current challenges
and suggest future directions.