title: Efficient Detection and Purification of Cell Populations Using Synthetic MicroRNA Switches.
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.
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Miki
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan; Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa-shi, Chiba, 277-8562, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Endo
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Seiya
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan; Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Funakoshi
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Ikue
authors.initials: I
authors.lastname: Takei
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Katayama
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Toyoda
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Maki
authors.initials: M
authors.lastname: Kotaka
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Takaki
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan; Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Umeda
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Okubo
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Misato
authors.initials: M
authors.lastname: Nishikawa
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Oishi
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Narita
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Ito
authors.initials: I
authors.lastname: Miyashita
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Asano
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Karin
authors.initials: K
authors.lastname: Hayashi
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Osafune
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto 606-8507, Japan; Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158, USA.
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamanaka
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan. Electronic address: hirohide.saito@cira.kyoto-u.ac.jp.
authors.forename: Hirohide
authors.initials: H
authors.lastname: Saito
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, Kyoto, 606-8507, Japan. Electronic address: yoshinor@cira.kyoto-u.ac.jp.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Yoshida
mesh_terms: D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001483:Base Sequence; D000072224:Bcl-2-Like Protein 11; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D006367:HeLa Cells; D022781:Hepatocytes / Q000166:cytology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology; D050417:Insulin-Secreting Cells / Q000166:cytology; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000637:transplantation; D009928:Organ Specificity; D011518:Proto-Oncogene Proteins / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; C000606776:Bcl2l11 protein, mouse; D008565:Membrane Proteins; D035683:MicroRNAs; D011518:Proto-Oncogene Proteins