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

Brian M. Wile, Kiwon Ban, Youngsup Yoon, Gang Bao

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Molecular 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 1/497% purity, as confirmed by electrophysiology and immunocytochemistry. After designing MBs, their ordering and validation requires 2 weeks, and the isolation process requires 3 h.

Original languageEnglish
Pages (from-to)2411-2424
Number of pages14
JournalNature Protocols
Volume9
Issue number10
DOIs
Publication statusPublished - 2014 Jan 1

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Stem cells
Purification
Cells
Pluripotent Stem Cells
Messenger RNA
Electrophysiology
Sorting
Oligonucleotides
RNA Transport
Therapeutic Human Experimentation
Cardiac Myocytes
Population
Immunohistochemistry

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Wile, Brian M. ; Ban, Kiwon ; Yoon, Youngsup ; Bao, Gang. / Molecular beacon-enabled purification of living cells by targeting cell type-specific mRNAs. In: Nature Protocols. 2014 ; Vol. 9, No. 10. pp. 2411-2424.
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Molecular beacon-enabled purification of living cells by targeting cell type-specific mRNAs. / Wile, Brian M.; Ban, Kiwon; Yoon, Youngsup; Bao, Gang.

In: Nature Protocols, Vol. 9, No. 10, 01.01.2014, p. 2411-2424.

Research output: Contribution to journalArticle

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