Global MicroRNA Analysis in Primary Hepatocyte Cultures.
MicroRNAs are small non-coding molecules that regulate gene expression and in return affect diverse biological functions, including those involved in toxicity and development of disease. Recent evidence suggests that microRNAs play an important role in liver pathologies, like viral hepatitis, alcoholic liver, hepatocellular carcinoma, or drug-induced liver injury. Furthermore, numerous studies demonstrated the high potential of microRNAs as promising non-invasive biomarkers of liver disease or as relevant targets for therapeutic treatment. This chapter describes a method for global microRNA analysis of primary hepatocytes by high-throughput sequencing. The method comprises the isolation of high-quality total RNA, analysis of microRNA sequencing data, and the validation of the findings by reverse transcriptase quantitative polymerase chain reaction analysis.
Faculty of Health, Medicine and Life Sciences, Department of Toxicogenomics, Maastricht University, Universiteitssingel 50, 6229ER, Maastricht, The Netherlands, j.krauskopf@maastrichtuniversity.nl.
Julian
J
Krauskopf
Almudena
A
Espín-Pérez
Jos C
JC
Kleinjans
Theo M
TM
de Kok
D019295:Computational Biology / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D061251:Primary Cell Culture / Q000379:methods; D015203:Reproducibility of Results
D035683:MicroRNAs