Next-Generation Sequencing to Investigate Urinary microRNAs from Macaca fascicularis (Cynomolgus Monkey)

Methods Mol Biol. 2017:1641:349-378. doi: 10.1007/978-1-4939-7172-5_20.

Abstract

Advanced sequencing technologies like next-generation sequencing (NGS) not only detect microRNAs (miRNAs) in biological samples but also facilitate de novo identification of miRNAs. Using an Ion Torrent's Ion Proton System, here we described miRNAs sequencing of urine samples collected from Macaca fascicularis (Cynomolgus monkey) to investigate miRNAs as potential novel biomarkers of nephrotoxicity in this species. Urinary miRNA sequencing methodologies described here include (a) urinary exosomal RNA isolation, (b) sequencing library preparation, (c) sequencing template preparation, and (d) template library sequencing using Ion Proton System. The sequencing method presented in this study serves as a valuable resource in the identification of novel urinary miRNAs in M. fascicularis.

Keywords: Biomarkers; Cynomolgus monkey; Ion Torrent; Kidney; Next-Generation Sequencing; Urine; microRNAs.

MeSH terms

  • Animals
  • Biomarkers / urine*
  • Gene Expression Profiling / methods
  • High-Throughput Nucleotide Sequencing / methods*
  • Kidney / metabolism
  • Macaca fascicularis
  • MicroRNAs / urine*
  • Sequence Analysis, RNA

Substances

  • Biomarkers
  • MicroRNAs