%0 Journal Article
%T Accurate RNA consensus sequencing for high-fidelity detection of transcriptional mutagenesis-induced epimutations.
%A Reid-Bayliss KS
%A Loeb LA
%J Proc Natl Acad Sci U S A
%V 114
%N 35
%D 08 2017 29
%M 28798064
%F 12.779
%R 10.1073/pnas.1709166114
%X 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.