| RRC ID |
48814
|
| Author |
Inagaki S, Tsutsui H, Suzuki K, Agetsuma M, Arai Y, Jinno Y, Bai G, Daniels MJ, Okamura Y, Matsuda T, Nagai T.
|
| Title |
Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging.
|
| Journal |
Sci Rep
|
| Abstract |
We report development of the first genetically encoded bioluminescent indicator for membrane voltage called LOTUS-V. Since it is bioluminescent, imaging LOTUS-V does not require external light illumination. This allows bidirectional optogenetic control of cellular activity triggered by Channelrhodopsin2 and Halorhodopsin during voltage imaging. The other advantage of LOTUS-V is the robustness of a signal-to-background ratio (SBR) wherever it expressed, even in the specimens where autofluorescence from environment severely interferes fluorescence imaging. Through imaging of moving cardiomyocyte aggregates, we demonstrated the advantages of LOTUS-V in long-term imaging are attributable to the absence of phototoxicity, and photobleaching in bioluminescent imaging, combined with the ratiometric aspect of LOTUS-V design. Collectively LOTUS-V extends the scope of excitable cell control and simultaneous voltage phenotyping, which should enable applications in bioscience, medicine and pharmacology previously not possible.
|
| Volume |
7
|
| Pages |
42398
|
| Published |
2017-2-13
|
| DOI |
10.1038/srep42398
|
| PII |
srep42398
|
| PMID |
28205521
|
| PMC |
PMC5322354
|
| MeSH |
Animals
Cell Line
Cells, Cultured
Electrophysiological Phenomena
Gene Expression*
Genes, Reporter*
Humans
Induced Pluripotent Stem Cells / metabolism
Kinetics
Luminescent Measurements* / methods
Luminescent Proteins / chemistry
Luminescent Proteins / genetics*
Luminescent Proteins / metabolism*
Models, Molecular
Molecular Imaging* / methods
Optogenetics* / methods
Protein Conformation
|
| IF |
3.998
|
| Times Cited |
25
|
|
WOS Category
|
NEUROSCIENCES
|
| Resource |
| Human and Animal Cells |
293T(RCB2202)
293(RCB1637) |
| DNA material |
CS-CDF-EG-PRE (RDB04380)
pCAG-HIVgp (RDB04394)
pCMV-VSV-G-RSV-Rev (RDB04393)
LOTUS-V/pcDNA3 (RDB19881) |
