Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches

Kenji Miki, Kei Endo, Seiya Takahashi, Shunsuke Funakoshi, Ikue Takei, Shota Katayama, Taro Toyoda, Maki Kotaka, Tadashi Takaki, Masayuki Umeda, Chikako Okubo, Misato Nishikawa, Akiko Oishi, Megumi Narita, Ito Miyashita, Kanako Asano, Karin Hayashi, Kenji Osafune, Shinya Yamanaka, Hirohide SaitoYoshinori Yoshida

Research output: Contribution to journalArticlepeer-review

101 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)699-711
Number of pages13
JournalCell Stem Cell
Volume16
Issue number6
DOIs
Publication statusPublished - 2015 Jun 4

ASJC Scopus subject areas

  • Molecular Medicine
  • Genetics
  • Cell Biology

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