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Molecular beacon-enabled purification of living cells by targeting cell type-specific mRNAs

DC FieldValueLanguage
dc.contributor.author윤영섭-
dc.date.accessioned2015-12-28T11:14:32Z-
dc.date.available2015-12-28T11:14:32Z-
dc.date.issued2014-
dc.identifier.issn1754-2189-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/139010-
dc.description.abstractMolecular beacons (MBs) are dual-labeled oligonucleotides that fluoresce only in the presence of complementary mRNA. The use of MBs to target specific mRNAs allows sorting of specific cells from a mixed cell population. In contrast to existing approaches that are limited by available surface markers or selectable metabolic characteristics, the MB-based method enables the isolation of a wide variety of cells. For example, the ability to purify specific cell types derived from pluripotent stem cells (PSCs) is important for basic research and therapeutics. In addition to providing a general protocol for MB design, validation and nucleofection into cells, we describe how to isolate a specific cell population from differentiating PSCs. By using this protocol, we have successfully isolated cardiomyocytes differentiated from mouse or human PSCs (hPSCs) with ∼ 97% purity, as confirmed by electrophysiology and immunocytochemistry. After designing MBs, their ordering and validation requires 2 weeks, and the isolation process requires 3 h.-
dc.description.statementOfResponsibilityopen-
dc.format.extent2411~2424-
dc.relation.isPartOfNATURE PROTOCOLS-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.subject.MESHAnimals-
dc.subject.MESHBiomarkers/metabolism-
dc.subject.MESHCell Differentiation-
dc.subject.MESHCell Separation/instrumentation-
dc.subject.MESHCell Separation/methods*-
dc.subject.MESHCells, Cultured-
dc.subject.MESHElectrophysiology/methods-
dc.subject.MESHFlow Cytometry/methods-
dc.subject.MESHHumans-
dc.subject.MESHMice-
dc.subject.MESHMolecular Biology/instrumentation-
dc.subject.MESHMolecular Biology/methods*-
dc.subject.MESHMolecular Probes/chemistry-
dc.subject.MESHMyocytes, Cardiac/cytology-
dc.subject.MESHOligonucleotides/chemistry-
dc.subject.MESHOligonucleotides/metabolism*-
dc.subject.MESHOrgan Specificity-
dc.subject.MESHPluripotent Stem Cells/cytology*-
dc.subject.MESHPluripotent Stem Cells/physiology-
dc.subject.MESHRNA, Messenger/analysis-
dc.subject.MESHRNA, Messenger/metabolism*-
dc.titleMolecular beacon-enabled purification of living cells by targeting cell type-specific mRNAs-
dc.typeArticle-
dc.contributor.collegeCollege of Medicine (의과대학)-
dc.contributor.departmentDept. of Life Science (의생명과학부)-
dc.contributor.googleauthorBrian M. Wile-
dc.contributor.googleauthorKiwon Ban-
dc.contributor.googleauthorYoung-Sup Yoon-
dc.contributor.googleauthorGang Bao-
dc.identifier.doi10.1038/nprot.2014.154-
dc.admin.authorfalse-
dc.admin.mappingfalse-
dc.contributor.localIdA02579-
dc.relation.journalcodeJ02300-
dc.identifier.eissn1750-2799-
dc.identifier.pmid25232937-
dc.contributor.alternativeNameYoon, Young Sup-
dc.contributor.affiliatedAuthorYoon, Young Sup-
dc.citation.volume9-
dc.citation.number10-
dc.citation.startPage2411-
dc.citation.endPage2424-
dc.identifier.bibliographicCitationNATURE PROTOCOLS, Vol.9(10) : 2411-2424, 2014-
Appears in Collections:
1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers

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