Department of Molecular Biophysics & Biochemistry, Yale University, New Haven, Connecticut, USA.
Chemical Biology Institute, Yale University, West Haven, Connecticut, USA.
Nat Methods. 2018 Mar;15(3):221-225. doi: 10.1038/nmeth.4582. Epub 2018 Jan 22.
RNA sequencing (RNA-seq) offers a snapshot of cellular RNA populations, but not temporal information about the sequenced RNA. Here we report TimeLapse-seq, which uses oxidative-nucleophilic-aromatic substitution to convert 4-thiouridine into cytidine analogs, yielding apparent U-to-C mutations that mark new transcripts upon sequencing. TimeLapse-seq is a single-molecule approach that is adaptable to many applications and reveals RNA dynamics and induced differential expression concealed in traditional RNA-seq.
RNA 测序 (RNA-seq) 提供了细胞 RNA 群体的快照，但没有关于测序 RNA 的时间信息。在这里，我们报告了 TimeLapse-seq，它使用氧化-亲核-芳香取代将 4-硫尿苷转化为胞苷类似物，在测序时产生新转录本的表观 U 到 C 突变。TimeLapse-seq 是一种单分子方法，适用于许多应用，揭示了传统 RNA-seq 中隐藏的 RNA 动态和诱导的差异表达。
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