Transposase-assisted tagmentation of RNA/DNA hybrid duplexes

• DOI: 10.7554/elife.54919
• Publication Date: 2020-07-23T12:10:12Z
• Authors: Bo Lu; Liting Dong; Danyang Yi; Meiling Zhang; Chenxu Zhu; Xiaoyu Li; Chengqi Yi
• Keywords: 0301 basic medicine; Artificial intelligence; Molecular biology; Genome Evolution and Polyploidy in Plants; Transposases; Plant Science; Gene; Agricultural and Biological Sciences; Computational biology; Transposition (logic); Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated proteins; epitranscriptome; RNA-Guided; DNA sequencing; RNA-Seq; Biology (General); Genome; Q; R; Life Sciences; Chromatin; RNA Methylation and Modification in Gene Expression; RNA methylation; Medicine; Genomic library; Sequence Analysis; Transposase; QH301-705.5; Science; Transposable element; 03 medical and health sciences; Biochemistry and Chemical Biology; Biochemistry, Genetics and Molecular Biology; Tn5; transposition; Genetics; Humans; Molecular Biology; Biology; Transposable Elements; tagmentation; DNA; RNA/DNA hybrids; Computer science; Base sequence; HEK293 Cells; FOS: Biological sciences; RNA; Gene expression; RNA-seq; Transcriptome; tRNA fragments

Tn5-mediated transposition of double-strand DNA has been widely utilized in various high-throughput sequencing applications. Here, we report that the Tn5 transposase is also capable of direct tagmentation of RNA/DNA hybrids in vitro. As a proof-of-concept application, we utilized this activity to replace the traditional library construction procedure of RNA sequencing, which contains many laborious and time-consuming processes. Results ofTransposase-assistedRNA/DNAhybridsCo-tagmEntation (termed ‘TRACE-seq’) are compared to traditional RNA-seq methods in terms of detected gene number, gene body coverage, gene expression measurement, library complexity, and differential expression analysis. At the meantime, TRACE-seq enables a cost-effective one-tube library construction protocol and hence is more rapid (within 6 hr) and convenient. We expect this tagmentation activity on RNA/DNA hybrids to have broad potentials on RNA biology and chromatin research.