Filter-Based Extracellular Vesicle mRNA Isolation and High-Throughput Gene Expression Analysis

Methods Mol Biol. 2017:1660:55-63. doi: 10.1007/978-1-4939-7253-1_6.

Abstract

Extracellular vesicles (EVs) are a heterogeneous group of membrane-encapsulated particles with different ranges of size, density, and cargo. Various types of RNA including mRNA are enclosed within EVs and can serve as novel biomarkers for disease detection and patient management. Ultracentrifugation, precipitation , antibody-based capture and filter-based methods are available as in-house laboratory procedures or commercially available kits to isolate EVs. Here, we describe a filter-based method for EV mRNA isolation that is designed for parallel processing of large sample numbers.

Keywords: Ascites; Exosomes; Extracellular vesicles; Filter; Gene expression; High-throughput; Isolation; Microvesicles; Urine; mRNA.

MeSH terms

  • Ascites / metabolism
  • Biomarkers
  • Exosomes
  • Extracellular Vesicles / metabolism*
  • Gene Expression Profiling / methods*
  • High-Throughput Screening Assays*
  • Humans
  • Micropore Filters*
  • Plasma / chemistry
  • RNA, Messenger / genetics*
  • RNA, Messenger / isolation & purification*
  • Specimen Handling / methods
  • Urine / chemistry

Substances

  • Biomarkers
  • RNA, Messenger