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Published on: March 20, 2021
DNA probes for monitoring dynamic and transient molecular encounters on live cell membranes
Mingxu You1,2,3,4, Yifan Lyu1,2, Da Han1,2
1Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha 410082, People's Republic of China.
Abstract:
Cells interact with the extracellular environment through molecules expressed on the membrane. Disruption of these membrane-bound interactions (or encounters) can result in disease progression. Advances in super-resolution microscopy have allowed membrane encounters to be examined, however, these methods cannot image entire membranes and cannot provide information on the dynamic interactions between membrane-bound molecules. Here, we show a novel DNA probe that can transduce transient membrane encounter events into readable cumulative fluorescence signals. The probe, which translocates from one anchor site to another, mimicking motor proteins, is realized through a toehold-mediated DNA strand displacement reaction. Using this probe, we successfully monitored rapid encounter events of membrane lipid domains using flow cytometry and fluorescence microscopy. Our results show a preference for encounters within the same lipid domains.
Insights
Researchers developed a novel DNA probe to track transient cell membrane encounters. This tool monitors rapid lipid domain interactions, revealing a preference for encounters within the same domain.
Area of Science:
- Cellular Biology
- Biochemistry
- Molecular Biology
Background:
- Cellular membranes are crucial for interacting with the extracellular environment.
- Disruptions in membrane-bound interactions are linked to disease progression.
- Current super-resolution microscopy methods have limitations in imaging dynamic membrane encounters.
Purpose of the Study:
- To develop a novel DNA probe for detecting transient membrane encounter events.
- To transduce these encounters into cumulative fluorescence signals for analysis.
- To overcome limitations of existing microscopy techniques for dynamic membrane studies.
Main Methods:
- Utilized a novel DNA probe based on toehold-mediated DNA strand displacement.
- Designed the probe to translocate between anchor sites, mimicking motor proteins.
- Employed flow cytometry and fluorescence microscopy to monitor membrane lipid domain encounters.
Main Results:
- Successfully monitored rapid encounter events of membrane lipid domains.
- The DNA probe effectively transduced transient encounters into cumulative fluorescence signals.
- Observed a preference for encounters occurring within the same lipid domains.
Conclusions:
- The novel DNA probe provides a new method for studying dynamic membrane interactions.
- This approach allows for the monitoring of rapid cellular encounters in real-time.
- Findings suggest specific lipid domain interactions play a role in cellular processes.
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