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Resolving the Unresolvable: Nanopore Sequencing as a Comprehensive Quality Control Platform for Gene Therapy Vectors - PubMed

5 hours ago
  • #nanopore sequencing
  • #gene therapy vectors
  • #quality control
  • Nanopore sequencing provides long-read, native single-molecule sequencing for quality control in gene therapy, addressing gaps left by traditional methods like Sanger and short-read NGS.
  • For plasmid DNA, it confirms full-length circular identity and detects structural heterogeneity, offering more detail than restriction mapping.
  • In viral vectors (AAV and lentivirus), it distinguishes full genomes from truncations and identifies sequence-resolved impurities, such as reverse-packaged plasmid backbones.
  • For mRNA therapeutics, direct RNA sequencing profiles poly(A) tail length distributions and base modifications (e.g., m1A) in a single assay.
  • The technology supports adaptive sampling for impurity enrichment and native epigenetic profiling, with potential to become a validated lot-release method despite current accuracy and compliance limitations.