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

Authors
Kwon Y.-S.Song H.
Issue Date
2016
Publisher
Taylor and Francis Ltd.
Keywords
AGO2; hESCs; microRNA
Citation
Animal Cells and Systems, v.20, no.1, pp 24 - 30
Pages
7
Indexed
SCIE
SCOPUS
KCI
Journal Title
Animal Cells and Systems
Volume
20
Number
1
Start Page
24
End Page
30
URI
https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/7052
DOI
10.1080/19768354.2015.1137227
ISSN
1976-8354
2151-2485
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. © 2016 Korean Society for Integrative Biology.
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