Veeranagouda Yaligara, Léonard Jean-François, Gautier Jean-Charles, Boitier Eric
Molecular Biology & Genomics, Translational Science, Sanofi R&D, Chilly-Mazarin, 91385, France.
Translational Medicine and Early Development, Biomarkers and Clinical Bioanalyses-Genomics, Sanofi R&D, Vitry-sur-Seine, France.
Methods Mol Biol. 2017;1641:349-378. doi: 10.1007/978-1-4939-7172-5_20.
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.
像新一代测序(NGS)这样的先进测序技术不仅能检测生物样本中的微小RNA(miRNA),还有助于miRNA的从头鉴定。在此,我们使用Ion Torrent的Ion Proton系统描述了从食蟹猴(猕猴)收集的尿液样本的miRNA测序,以研究miRNA作为该物种肾毒性潜在新生物标志物的情况。这里描述的尿液miRNA测序方法包括:(a)尿液外泌体RNA分离,(b)测序文库制备,(c)测序模板制备,以及(d)使用Ion Proton系统进行模板文库测序。本研究中提出的测序方法是鉴定食蟹猴新尿液miRNA的宝贵资源。