Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches

Cell Stem Cell. 2015 Jun 4;16(6):699-711. doi: 10.1016/j.stem.2015.04.005. Epub 2015 May 21.


Isolation of specific cell types, including pluripotent stem cell (PSC)-derived populations, is frequently accomplished using cell surface antigens expressed by the cells of interest. However, specific antigens for many cell types have not been identified, making their isolation difficult. Here, we describe an efficient method for purifying cells based on endogenous miRNA activity. We designed synthetic mRNAs encoding a fluorescent protein tagged with sequences targeted by miRNAs expressed by the cells of interest. These miRNA switches control their translation levels by sensing miRNA activities. Several miRNA switches (miR-1-, miR-208a-, and miR-499a-5p-switches) efficiently purified cardiomyocytes differentiated from human PSCs, and switches encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as miR-126-, miR-122-5p-, and miR-375-switches purified endothelial cells, hepatocytes, and insulin-producing cells differentiated from hPSCs, respectively. Thus, miRNA switches can purify cell populations for which other isolation strategies are unavailable.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Apoptosis Regulatory Proteins / metabolism
  • Base Sequence
  • Bcl-2-Like Protein 11
  • Cell Separation / methods*
  • Flow Cytometry
  • HeLa Cells
  • Hepatocytes / cytology
  • Human Umbilical Vein Endothelial Cells / cytology
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Insulin-Secreting Cells / cytology
  • Membrane Proteins / metabolism
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Myocardium / cytology
  • Myocardium / metabolism
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / transplantation
  • Organ Specificity
  • Proto-Oncogene Proteins / metabolism


  • Apoptosis Regulatory Proteins
  • BCL2L11 protein, human
  • Bcl-2-Like Protein 11
  • Bcl2l11 protein, mouse
  • Membrane Proteins
  • MicroRNAs
  • Proto-Oncogene Proteins

Associated data

  • GEO/GSE60633