Fast and Simplified Method for High Through-put Isolation of miRNA from Highly Purified High Density Lipoprotein

• Published in: Journal of visualized experiments 2016-07 (113)
• Main Authors: Seneshaw, Mulugeta, Mirshahi, Faridoddin, Min, Hae-Ki, Asgharpour, Amon, Mirshahi, Shervin, Daita, Kalyani, Boyett, Sherry, Santhekadur, Prasanna K, Fuchs, Michael, Sanyal, Arun J
• Format: Article
• Language: English
• Subjects: Biological Transport; Biomarkers; Electrophoresis, Agar Gel; Humans; Lipoproteins, HDL - chemistry; Lipoproteins, HDL - isolation & purification; MicroRNAs - isolation & purification; Molecular Biology; Prognosis; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/54257

Small non-coding RNAs (miRNAs) have been implicated in a variety of human diseases including metabolic syndromes. They may be utilized as biomarkers for diagnosis and prognosis or may serve as targets for drug development, respectively. Recently it has been shown that miRNAs are carried in lipoproteins, particularly high density lipoproteins (HDL) and are delivered to recipient cells for uptake. This raises the possibility that miRNAs play a critical and pivotal role in cellular and organ function via regulation of gene expression as well as messenger for cell-cell communications and crosstalk between organs. Current methods for miRNA isolation from purified HDL are impractical when utilizing small samples on a large scale. This is largely due to the time consuming and laborious methods used for lipoprotein isolation. We have developed a simplified approach to rapidly isolate purified HDL suitable for miRNA analysis from plasma samples. This method should facilitate investigations into the role of miRNAs in health and disease and in particular provide new insights into the variety of biological functions, outside of the reverse cholesterol transport, that have been ascribed to HDL. Also, the miRNA species which are present in HDL can provide valuable information of clinical biomarkers for diagnosis of various diseases.