Novel microRNA discovery using small RNA sequencing in post-mortem human brain

• Published in: BMC genomics Vol. 17; no. 1; p. 776
• Main Authors: Wake, Christian, Labadorf, Adam, Dumitriu, Alexandra, Hoss, Andrew G, Bregu, Joli, Albrecht, Kenneth H, DeStefano, Anita L, Myers, Richard H
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 04.10.2016; BioMed Central
• Subjects: Autopsy; Brain - metabolism; Brain - pathology; Brain research; Gene Expression Profiling; Gene Expression Regulation; High-Throughput Nucleotide Sequencing; Humans; Huntington Disease - genetics; MicroRNA; MicroRNAs - genetics; Nervous system diseases; Parkinson Disease - genetics; RNA sequencing; Novel miRNA discovery; Neurodegenerative disease; MicroRNA; Prefrontal cortex; miRNA sequencing
• ISSN: 1471-2164; 1471-2164
• DOI: 10.1186/s12864-016-3114-3

MicroRNAs (miRNAs) are short, non-coding RNAs that regulate gene expression mainly through translational repression of target mRNA molecules. More than 2700 human miRNAs have been identified and some are known to be associated with disease phenotypes and to display tissue-specific patterns of expression. We used high-throughput small RNA sequencing to discover novel miRNAs in 93 human post-mortem prefrontal cortex samples from individuals with Huntington's disease (n = 28) or Parkinson's disease (n = 29) and controls without neurological impairment (n = 36). A custom miRNA identification analysis pipeline was built, which utilizes miRDeep* miRNA identification and result filtering based on false positive rate estimates. Ninety-nine novel miRNA candidates with a false positive rate of less than 5 % were identified. Thirty-four of the candidate miRNAs show sequence similarity with known mature miRNA sequences and may be novel members of known miRNA families, while the remaining 65 may constitute previously undiscovered families of miRNAs. Nineteen of the 99 candidate miRNAs were replicated using independent, publicly-available human brain RNA-sequencing samples, and seven were experimentally validated using qPCR. We have used small RNA sequencing to identify 99 putative novel miRNAs that are present in human brain samples.