Analysis of microRNAs in a knock-in hESC line expressing epitope-tagged AGO2

Young Soo Kwon, Hoseok Song

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Argonaute (AGO) family proteins, which are key components of RNA-induced silencing complexes (RISCs), associate with both microRNAs (miRNAs) and mRNAs to regulate gene expression at post-transcriptional stages. While high-quality antibodies for AGO proteins are extremely useful for studies on the functions of miRNAs and AGO proteins, they are not always readily available. In this study, we generated a knock-in human embryonic stem cell (hESC) line by using homologous recombination to express FLAG-tagged AGO2 from its native genomic locus. FLAG-tagged AGO2 was exploited to analyze AGO2-bound miRNAs in hESCs cultured on a feeder layer by immunoprecipitation. Furthermore, the pluripotency of ESCs allowed for the analysis of miRNAs after differentiation into fibroblasts and cardiomyocytes. The genetically modified hESC line generated in this study will be highly useful for studying the functions of miRNA-mediated regulation in human development and cell differentiation.

Original languageEnglish
Pages (from-to)24-30
Number of pages7
JournalAnimal Cells and Systems
Volume20
Issue number1
DOIs
Publication statusPublished - 2016 Jan 2

Fingerprint

embryonic stem cells
Stem cells
MicroRNAs
microRNA
Argonaute Proteins
epitopes
Epitopes
cell lines
Cell Line
RNA-Induced Silencing Complex
Feeder Cells
proteins
human development
Homologous Recombination
homologous recombination
Human Development
Fibroblasts
Immunoprecipitation
Cardiac Myocytes
Gene expression

Keywords

  • AGO2
  • hESCs
  • microRNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Animal Science and Zoology

Cite this

Analysis of microRNAs in a knock-in hESC line expressing epitope-tagged AGO2. / Kwon, Young Soo; Song, Hoseok.

In: Animal Cells and Systems, Vol. 20, No. 1, 02.01.2016, p. 24-30.

Research output: Contribution to journalArticle

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