| Chapter title |
Visualizing RNA in Live Bacterial Cells Using Fluorophore- and Quencher-Binding Aptamers
|
|---|---|
| Chapter number | 19 |
| Book title |
RNA Detection
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7213-5_19 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7212-8, 978-1-4939-7213-5
|
| Authors |
Murat Sunbul, Ankita Arora, Andres Jäschke |
| Abstract |
To elucidate the roles, dynamics, and regulation of RNAs, it is vital to be able to visualize the RNA of interest (ROI) in living cells noninvasively. Here, we describe a novel live-cell RNA imaging method using fluorophore- and quencher-binding aptamers, which can be genetically fused to the ROI. In this method, new membrane permeable and nonfluorescent fluorophore-quencher conjugates were utilized, and we showed that their fluorescence increases dramatically upon binding to fluorophore- or quencher-binding aptamers. This phenomenon allowed for labeling the ROI with many different colored fluorophores and also dual-color imaging of two distinct RNAs in live bacteria. Our approach uses small RNA tags and small molecule fluorophores for labeling, thereby minimal perturbation on the function and dynamics of the RNA of interest is expected. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 11 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 36% |
| Professor | 1 | 9% |
| Unspecified | 1 | 9% |
| Researcher | 1 | 9% |
| Other | 1 | 9% |
| Other | 0 | 0% |
| Unknown | 3 | 27% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 4 | 36% |
| Biochemistry, Genetics and Molecular Biology | 2 | 18% |
| Agricultural and Biological Sciences | 1 | 9% |
| Unspecified | 1 | 9% |
| Unknown | 3 | 27% |