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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Watanabe, Tomonobu M. Takai, Akira Saito, Kenta Okada, Yasushi Nagai, Takeharu Ohyanagi, Tatsuya Nakano, Masahiro Haruno, Remi Jin, Takashi |
| Description | Author Affiliation: Takai A ( Laboratories for Cell Polarity Regulation.); Nakano M ( The Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan); Saito K ( The Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan); Haruno R ( The Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan); Watanabe TM ( Comprehensive Bioimaging, PRESTO, Japan Science and Technology Agency, Tokyo 102-0075, Japan.); Ohyanagi T ( Nano-Bio Probes, and.); Jin T ( Nano-Bio Probes, and.); Okada Y ( Laboratories for Cell Polarity Regulation, ng1@sanken.osaka-u.ac.jp y.okada@riken.jp.); Nagai T ( The Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047, Japan); |
| Abstract | Fluorescence live imaging has become an essential methodology in modern cell biology. However, fluorescence requires excitation light, which can sometimes cause potential problems, such as autofluorescence, phototoxicity, and photobleaching. Furthermore, combined with recent optogenetic tools, the light illumination can trigger their unintended activation. Because luminescence imaging does not require excitation light, it is a good candidate as an alternative imaging modality to circumvent these problems. The application of luminescence imaging, however, has been limited by the two drawbacks of existing luminescent protein probes, such as luciferases: namely, low brightness and poor color variants. Here, we report the development of bright cyan and orange luminescent proteins by extending our previous development of the bright yellowish-green luminescent protein Nano-lantern. The color change and the enhancement of brightness were both achieved by bioluminescence resonance energy transfer (BRET) from enhanced Renilla luciferase to a fluorescent protein. The brightness of these cyan and orange Nano-lanterns was ∼20 times brighter than wild-type Renilla luciferase, which allowed us to perform multicolor live imaging of intracellular submicron structures. The rapid dynamics of endosomes and peroxisomes were visualized at around 1-s temporal resolution, and the slow dynamics of focal adhesions were continuously imaged for longer than a few hours without photobleaching or photodamage. In addition, we extended the application of these multicolor Nano-lanterns to simultaneous monitoring of multiple gene expression or $Ca^{2+}$ dynamics in different cellular compartments in a single cell. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 14 |
| Volume Number | 112 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2015-04-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Luciferases Chemistry Luminescence Luminescent Proteins Recombinant Fusion Proteins Animals Calcium Metabolism Cell Line DNA Embryonic Stem Cells Cytology Endosomes Fluorescence Resonance Energy Transfer Focal Adhesions Gene Expression Regulation Luciferases, Renilla Mice Molecular Sequence Data Oligonucleotides Peroxisomes Promoter Regions, Genetic Renilla Vinculin Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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