Dual optical biosensors for imaging microRNA-1 during myogenesis.
Dual optical microRNA (miRNA) imaging systems, bioluminescent reporter gene (a signal-off mechanism)- or fluorescent molecular beacon (MB) (a signal-on mechanism)-based miRNA imaging system, have individually allowed us to sense miRNA biogenesis in a noninvasive and iterative manner. Both of these imaging systems have shortcomings with respect to image quality. Therefore, we designed a dual optical imaging systems by simultaneous imaging of a miRNA-1 reporter gene (CMV/Gluc/3xPT_miR-1) and miRNA-1 MB in a single cell to overcome these limitations and used it to visualize miRNA-1, a highly expressed miRNA in cardiac and skeletal muscle. During myogenic differentiation of C2C12 cells, the bioluminescence intensity from CMV/Gluc/3xPT_miR-1 revealed a miRNA-1-dependent gradual decrease and the fluorescence intensity from miR-1 MB demonstrated a miRNA-1-dependent gradual increase both in vitro and in vivo. The dual miRNA-1 imaging systems, which provides the complementary imaging information about miRNA biogenesis, could be useful to sense miRNA expression during various biologic processes.
Division of Nuclear Medicine, Department of Radiology, Yonsei University College of Medicine, South Korea.
Won Jun
WJ
Kang
Ye Lim
YL
Cho
Ju Ri
JR
Chae
Jong Doo
JD
Lee
Bahy A
BA
Ali
Abdulaziz A
AA
Al-Khedhairy
Chang Hyun
CH
Lee
Soonhag
S
Kim
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D003952:Diagnostic Imaging / Q000379:methods*; D005453:Fluorescence; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D015335:Molecular Probes / Q000378:metabolism; D024510:Muscle Development; D055096:Optical Devices; D015203:Reproducibility of Results
D035683:MicroRNAs; D015335:Molecular Probes