Toehold-initiated rolling circle amplification for visualizing individual microRNAs in situ in single cells.
The ability to quantitate and visualize microRNAs (miRNAs) in situ in single cells would greatly facilitate the elucidation of miRNA-mediated regulatory circuits and their disease associations. A toehold-initiated strand-displacement process was used to initiate rolling circle amplification of specific miRNAs, an approach that achieves both stringent recognition and in situ amplification of the target miRNA. This assay, termed toehold-initiated rolling circle amplification (TIRCA), can be utilized to identify miRNAs at physiological temperature with high specificity and to visualize individual miRNAs in situ in single cells within 3 h. TIRCA is a competitive candidate technique for in situ miRNA imaging and may help us to understand the role of miRNAs in cellular processes and human diseases in more detail.
Department of Chemistry, Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Tsinghua University, Beijing 100084 (China).
Ruijie
R
Deng
Longhua
L
Tang
Qianqian
Q
Tian
Ying
Y
Wang
Lei
L
Lin
Jinghong
J
Li
D001483:Base Sequence; D002460:Cell Line; D006801:Humans; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D018613:Microscopy, Confocal / Q000379:methods; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D059010:Single-Cell Analysis / Q000379:methods*
D035683:MicroRNAs
RNA structures; in situ imaging; isothermal amplification; microRNAs; strand displacement