High-throughput characterization of primary microRNA transcripts

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Scopus citations

Abstract

Proper control of microRNA (miRNA) expression is critical for normal development and physiology, while abnormal miRNA expression is a common feature of many diseases. Dissecting mechanisms of miRNA regulation, however, is complicated by the generally poor annotation of miRNA primary transcripts (pri-miRNAs). Although some miRNAs are processed from well-defined protein coding genes, the majority of pri-miRNAs are poorly characterized noncoding RNAs, with incomplete annotation of promoters, splice sites, and polyadenylation signals. Due to the efficiency of DROSHA processing, the abundance of pri-miRNAs is very low at steady state, thereby complicating the elucidation of pri-miRNA structures. Here we describe a strategy to enrich intact pri-miRNAs and improve their coverage in RNA sequencing (RNA-seq) experiments. In addition, we outline a computational approach for reconstruction of pri-miRNA structures. This pipeline begins with raw RNA-seq reads and concludes with publication-ready visualization of pri-miRNA annotations. Together, these approaches allow the user to define and explore miRNA gene structures in a cell-type or organism of interest.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages1-9
Number of pages9
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1823
ISSN (Print)1064-3745

Keywords

  • Primary transcript
  • RNA-seq
  • Transcriptome assembly
  • microRNA
  • pri-miRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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