Chapter title |
Molecular Beacon-Based MicroRNA Imaging During Neurogenesis.
|
---|---|
Chapter number | 11 |
Book title |
RNA Imaging
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3148-4_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3147-7, 978-1-4939-3148-4
|
Authors |
Lee, Jonghwan, Kim, Soonhag, Jonghwan Lee, Soonhag Kim Ph.D., Soonhag Kim |
Editors |
Zdravka Medarova |
Abstract |
The fluorescence monitoring system for examining endogenous microRNA (miRNA) activity in cellular level provides crucial information on not only understanding a critical role of miRNA involving a variety of biological processes, but also evaluating miRNA expression patterns in a noninvasive manner. In this protocol, we report the details of a new procedure for a molecular beacon-based miRNA monitoring system, which includes the illustration scheme for miRNA detection strategy, exogenous miRNA detection, and measurement of endogenous miRNA expression level during neurogenesis. The fluorescence signal of miR-124a beacon quenched by BHQ2 was gradually recovered as increasing concentration of the miR-124a in tube. The functional work of miR-124a beacon was examined in intracellular environment, allowing for the internalization of the miR-124a beacon by lipofectamine, which resulted in activated fluorescent signals of the miR-124a beacon in the HeLa cells after the addition of synthetic miR-124a. The endogenous miR-124a expression level was detected by miR-124a beacon system during neurogenesis, showing brighter fluorescence intensity in cytoplasmic area of P19 cells after induction of neuronal differentiation by retinoic acid. The molecular beacon based-miRNA detection technique could be applicable to the simultaneous visualization of a variety of miRNA expression patterns using different fluorescence dyes. For the study of examining endogenous miRNA expression level using miRNA-beacon system, if cellular differentiation step is already prepared, transfection step of miR-124a beacon into P19 cells, and acquisition of activated fluorescence signal measured by confocal microscope can be conducted approximately within 6 h. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 2 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 7 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 2 | 29% |
Researcher | 2 | 29% |
Professor | 1 | 14% |
Student > Bachelor | 1 | 14% |
Professor > Associate Professor | 1 | 14% |
Other | 0 | 0% |
Readers by discipline | Count | As % |
---|---|---|
Neuroscience | 3 | 43% |
Agricultural and Biological Sciences | 2 | 29% |
Biochemistry, Genetics and Molecular Biology | 2 | 29% |