A color-tunable molecular beacon to sense miRNA-9 expression during neurogenesis.
A typical molecular beacon (MB) composing of a fluorophore and a quencher has been used to sense various intracellular biomolecules including microRNAs (miRNA, miR). However, the on/off-tunable miRNA MB is difficult to distinguish whether the observed low fluorescence brightness results from low miRNA expression or low transfection of the miRNA MB. We developed a color-tunable miRNA-9 MB (ColoR9 MB) to sense miR-9 expression-dependent color change. The ColoR9 MB was synthesized by a partially double-stranded DNA oligonucleotide containing a miR-9 binding site and a reporter probe with Cy3/black hole quencher 1 (BHQ1) at one end and a reference probe with Cy5.5 at the other end. The ColoR9 MB visualized CHO and P19 cells with red color in the absence of miR-9 and yellow color in the presence of miR-9. In vivo imaging demonstrated that the green fluorescence recovery of the reporter probe from the ColoR9 MB increased gradually during neuronal differentiation of P19 cells, whereas red fluorescence activity of the reference probe remained constant. These results showed the great specificity of sensing miR-9 expression- and neurogenesis-dependent color change.
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Hae Young
HY
Ko
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Jonghwan
J
Lee
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Jin Young
JY
Joo
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Yong Seung
YS
Lee
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Hyejung
H
Heo
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Jung Jae
JJ
Ko
Department of Biomedical Science, College of Life Science, CHA University, 605-21 Yeoksam 1-dong, Gangnam-gu, Seoul 135-081, Republic of Korea.
Soonhag
S
Kim
D000818:Animals; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D045744:Cell Line, Tumor; D003412:Cricetulus; D004247:DNA / Q000235:genetics; D005456:Fluorescent Dyes; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*; D055495:Neurogenesis / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000235:genetics*; D061848:Optical Imaging; D012097:Repressor Proteins / Q000235:genetics; D000099229:RE1-Silencing Transcription Factor
D004247:DNA; D005456:Fluorescent Dyes; D035683:MicroRNAs; D015345:Oligonucleotide Probes; D012097:Repressor Proteins; D000099229:RE1-Silencing Transcription Factor; C551804:MIRN9 microRNA, mouse