All-in-One Synchronized DNA Nanodevices Facilitating Multiplexed Cell Imaging.

Area of Science:

• Nanotechnology
• Molecular Biology
• Biomedical Engineering

Background:

• Multifunctional DNA nanodevices are crucial for in vitro diagnostics and in situ cell imaging.
• Existing nanodevices often use split building blocks, leading to asynchronous behavior and loss of information in cellular environments.
• Challenges include inhomogeneous distribution, diffusion limitations, and complex post-assembly processes.

Purpose of the Study:

• To develop all-in-one DNA nanodevices for synchronous multiplexed imaging within cells.
• To overcome the limitations of split DNA nanodevices in complex cellular microenvironments.
• To enable accurate and efficient detection of intracellular targets.

Main Methods:

• Constructed all-in-one DNA nanodevices with all components modified on individual gold nanoparticles.
• Utilized target-specific walkers (hairpin tracks with damaged bases) for molecular recognition.
• Employed intracellular enzymes to actuate synchronized module motion after target binding.
• Applied nanodevices for multiplexed imaging of intracellular microRNA-21 and telomerase.

Main Results:

• Achieved integrated internalization of nanodevices into cells, enhancing local concentrations.
• Circumvented inhomogeneous distribution and diffusion limitations in the cytoplasm.
• Demonstrated synchronized, multiplexed cell imaging through enzyme-actuated on-particle module motion.
• Successfully monitored intracellular microRNA-21 and telomerase expression levels.

Conclusions:

• The developed all-in-one DNA nanodevices enable synchronous multiplexed imaging with high efficiency.
• This design overcomes key limitations of previous split-component nanodevices.
• The flexible platform can be adapted for detecting various cytoplasmic molecules and monitoring cellular pathways.