Journal
CELL
Volume 184, Issue 25, Pages 6193-+Publisher
CELL PRESS
DOI: 10.1016/j.cell.2021.11.005
Keywords
-
Categories
Funding
- NIH [S10OD016374, S10OD023548, K22CA212060, R01GM136711]
- Cervical Cancer SPORE [P50CA098252]
- W.W. Smith Charitable Trust [C1901]
- Sol Goldman Pancreatic Cancer Research Center
Ask authors/readers for more resources
Researchers have developed a set of barcoding proteins capable of generating over 100 barcodes, overcoming the limited multiplexing capacity of genetically encoded fluorescent biosensors in live cells. Simultaneous tracking of multiple biosensors reveals highly coordinated activities and complex interactions within the signaling network.
Genetically encoded fluorescent biosensors are powerful tools for monitoring biochemical activities in live cells, but their multiplexing capacity is limited by the available spectral space. We overcome this problem by developing a set of barcoding proteins that can generate over 100 barcodes and are spectrally separable from commonly used biosensors. Mixtures of barcoded cells expressing different biosensors are simultaneously imaged and analyzed by deep learning models to achieve massively multiplexed tracking of signaling events. Importantly, different biosensors in cell mixtures show highly coordinated activities, thus facilitating the delineation of their temporal relationship. Simultaneous tracking of multiple biosensors in the receptor tyrosine kinase signaling network reveals distinct mechanisms of effector adaptation, cell autonomous and non-autonomous effects of KRAS mutations, as well as complex interactions in the network. Biosensor barcoding presents a scalable method to expand multiplexing capabilities for deciphering the complexity of signaling networks and their interactions between cells.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available