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Title: Nanopore dwell time analysis permits sequencing and conformational assignment of pseudouridine in SARS-CoV-2
Authors: Fleming, Aaron M.
Keywords: Nanopore; giải trình tự; SARS-CoV-2; cấu trúc; pseudouridine
Issue Date: 2021
Publisher: bioRxiv
Abstract: Nanopore devices can directly sequence RNA, and the method has the potential to determine locations of epitranscriptomic modifications that have grown in significance because of their roles in cell regulation and stress response. Pseudouridine (Ψ), the most common modification in RNA, was sequenced with a nanopore system using a protein sensor with a helicase brake in synthetic RNAs with 100% modification at 18 known human pseudouridinylation sites. The new signals were compared to native uridine (U) control strands to characterize base calling and associated errors as well as ion current and dwell time changes. The data point to strong sequence context effects in which Ψ can easily be detected in some contexts while in others Ψ yields signals similar to U that would be false negatives in an unknown sample.
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