High-resolution Identification and Separation of Living Cell Types by Multiple microRNA-responsive Synthetic mRNAs

Sci Rep. 2016 Feb 23;6:21991. doi: 10.1038/srep21991.

Abstract

The precise identification and separation of living cell types is critical to both study cell function and prepare cells for medical applications. However, intracellular information to distinguish live cells remains largely inaccessible. Here, we develop a method for high-resolution identification and separation of cell types by quantifying multiple microRNA (miRNA) activities in live cell populations. We found that a set of miRNA-responsive, in vitro synthesized mRNAs identify a specific cell population as a sharp peak and clearly separate different cell types based on less than two-fold differences in miRNA activities. Increasing the number of miRNA-responsive mRNAs enhanced the capability for cell identification and separation, as we precisely and simultaneously distinguished different cell types with similar miRNA profiles. In addition, the set of synthetic mRNAs separated HeLa cells into subgroups, uncovering heterogeneity of the cells and the level of resolution achievable. Our method could identify target live cells and improve the efficiency of cell purification from heterogeneous populations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Separation / methods*
  • Gene Expression Regulation
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics*
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • MCF-7 Cells
  • MicroRNAs / chemical synthesis
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Plasmids / chemistry
  • Plasmids / metabolism
  • RNA, Messenger / chemical synthesis
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Transfection

Substances

  • Luminescent Proteins
  • MicroRNAs
  • RNA, Messenger
  • enhanced green fluorescent protein
  • red fluorescent protein
  • Green Fluorescent Proteins