PDF(Sci-hub)
Abstract：
Transcriptional mutagenesis (TM) due to misincorporation during RNA transcription can result in mutant RNAs, or epimutations, that generate proteins with altered properties. TM has long been hypothesized to play a role in aging, cancer, and viral and bacterial evolution. However, inadequate methodologies have limited progress in elucidating a causal association. We present a high-throughput, highly accurate RNA sequencing method to measure epimutations with single-molecule sensitivity. Accurate RNA consensus sequencing (ARC-seq) uniquely combines RNA barcoding and generation of multiple cDNA copies per RNA molecule to eliminate errors introduced during cDNA synthesis, PCR, and sequencing. The stringency of ARC-seq can be scaled to accommodate the quality of input RNAs. We apply ARC-seq to directly assess transcriptome-wide epimutations resulting from RNA polymerase mutants and oxidative stress.
摘要：
由于RNA转录过程中的错误掺入而导致的转录诱变（TM）可导致突变的RNA，或模仿，产生属性改变的蛋白质。长期以来，人们一直认为TM在衰老中起作用，癌症,以及病毒和细菌的进化。然而,不充分的方法限制了阐明因果关系的进展。我们提出了一个高通量，高精度的RNA测序方法，以单分子灵敏度测量表象。准确的RNA共有测序（ARC-seq）独特地结合了RNA条形码和每个RNA分子的多个cDNA拷贝的产生，以消除在cDNA合成过程中引入的错误。PCR,和测序。可以缩放ARC-seq的严格性以适应输入RNA的质量。我们应用ARC-seq直接评估由RNA聚合酶突变体和氧化应激引起的转录组范围的表观突变。
