Fast and Simplified Method for High Through-put Isolation of miRNA from Highly Purified High Density Lipoprotein. J Vis Exp 2016 Jul 27(113)
Date
08/09/2016Pubmed ID
27501005Pubmed Central ID
PMC5015653DOI
10.3791/54257Scopus ID
2-s2.0-84979918000 (requires institutional sign-in at Scopus site) 2 CitationsAbstract
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.
Author List
Seneshaw M, Mirshahi F, Min HK, Asgharpour A, Mirshahi S, Daita K, Boyett S, Santhekadur PK, Fuchs M, Sanyal AJAuthor
Shervin Mirshahi MD Assistant Professor in the Radiology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Biological TransportBiomarkers
Electrophoresis, Agar Gel
Humans
Lipoproteins, HDL
MicroRNAs
Prognosis
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
