Molecular beacon-based bioimaging of multiple microRNAs during myogenesis.
MicroRNAs (miRNAs, miR) are associated with multiple cellular processes and diseases. Here, we designed fluorescent DNA probes composed of stem loop-structured DNA complementary to miRNAs and fluorophore-quencher pairs [molecular beacon (MB)] to simultaneously monitor the biogenesis of miR-206 and miR-26a, which are highly expressed during myogenic differentiation. C2C12 cells were transfected with an MB targeting miR-26a and containing a 6-FAM-BHQ1 pair (miRNA-26a MB) or an MB targeting miR-206 with a Texas Red-BHQ2 pair (miRNA-206 MB). In vitro and in vivo fluorescence analysis revealed that, only in differentiated single C2C12 cell, significantly increased fluorescence signals of miRNA-26a MB, miRNA-206 MB or simultaneous incubation of both beacons were detected due to the hybridization of miR-206 or miR-26a with their respective beacons, resulting in activation of fluorescence. Our MB-based miRNA imaging system may serve as a new imaging probe for monitoring multiple miRNAs during various cellular or disease processes associated with miRNAs.
Division of Nuclear Medicine, Department of Nuclear Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea.
Won Jun
WJ
Kang
Ye Lim
YL
Cho
Ju Ri
JR
Chae
Jong Doo
JD
Lee
Kyung-Ju
KJ
Choi
Soonhag
S
Kim
D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D018613:Microscopy, Confocal; D024510:Muscle Development / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D035683:MicroRNAs