Chapter title |
Sensing miRNA: Signal Amplification by Cognate RISC for Intracellular Detection of miRNA in Live Cells.
|
---|---|
Chapter number | 10 |
Book title |
RNA Imaging
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3148-4_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3147-7, 978-1-4939-3148-4
|
Authors |
Kavishwar, Amol, Medarova, Zdravka, Amol Kavishwar, Zdravka Medarova |
Editors |
Zdravka Medarova |
Abstract |
The ability to detect miRNA expression in live cells would leave these cells available for further manipulation or culture. Here, we describe the design of a miRNA sensor oligonucleotide whose sequence mimics the target mRNA. The sensor has a fluorescent label on one end of the oligo and a quencher on the other. When inside the cell, the sensor is recognized by its cognate miRNA-RISC and gets cleaved, setting the fluorophore free from its quencher. This results in fluorescence "turn on." Since cleavage by the RISC complex is an enzymatic process, the described approach has a very high level of sensitivity (nM). The rate of nonspecific cleavage of the sensor is very slow permitting the collection of meaningful signal over a long period of time. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 4 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 2 | 50% |
Professor > Associate Professor | 1 | 25% |
Student > Ph. D. Student | 1 | 25% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 2 | 50% |
Agricultural and Biological Sciences | 1 | 25% |
Neuroscience | 1 | 25% |