RRC ID 38542
Author Iwagawa T, Ohuchi SP, Watanabe S, Nakamura Y.
Title Selection of RNA aptamers against mouse embryonic stem cells.
Journal Biochimie
Abstract Embryonic stem cells (ESCs) are capable of unlimited self-renewal and differentiation into multiple cell types. Recent large-scale analyses have identified various cell surface molecules on ESCs. Some of them are considered to be beneficial markers for characterization of cellular phenotypes and/or play an essential role for regulating the differentiation state. Thus, it is desired to efficiently produce affinity reagents specific to these molecules. In this study, to develop such reagents for mouse ESCs (mESCs), we selected RNA aptamers against intact, live mESCs using several selection strategies. The initial selection provided us with several anti-mESC aptamers of distinct sequences, which unexpectedly react with the same molecule on mESCs. Then, to isolate aptamers against different surface markers on mESCs, one of the selected aptamers was used as a competitor in the subsequent selections. In addition, one of the selections further employed negative selection against differentiated mouse cells. Consequently, we successfully isolated three classes of anti-mESC aptamers that do not compete with one another. The isolated aptamers were shown to distinguish mESCs from differentiated mouse cell lines and trace the differentiation process of mESCs. These aptamers could prove useful for developing molecular probes and manipulation tools for mESCs.
Volume 94(1)
Pages 250-7
Published 2012-1-1
DOI 10.1016/j.biochi.2011.10.017
PII S0300-9084(11)00413-5
PMID 22085640
MeSH Animals Aptamers, Nucleotide* Base Sequence Cells, Cultured DNA Primers Embryonic Stem Cells / cytology* Mice Molecular Sequence Data Sequence Homology, Nucleic Acid
IF 3.413
Times Cited 11
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells C2C12(RCB0987) Hepa 1-6(RCB1638) NB2a(RCB2639)