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A molecular delivery system by using AFM and nanoneedle.
Sung Woong Han1, Chikashi Nakamura, Ikuo Obataya
1Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakoji, Amagasaki, Hyogo 661-0974, Japan.
Biosensors & Bioelectronics
|March 3, 2005
Summary
Researchers created a novel system for low-invasive gene transfer into single cells. This atomic force microscope-nanoneedle system successfully delivered DNA into cells, demonstrating its potential for precise molecular manipulation.
Area of Science:
- Biotechnology
- Cell Biology
- Nanotechnology
Background:
- Precise manipulation of intracellular components is crucial for biological research and therapeutic applications.
- Existing methods for gene and molecule transfer can be invasive or lack precision at the single-cell level.
Purpose of the Study:
- To develop a novel, low-invasive system for manipulating and transferring genes or molecules within single living cells.
- To utilize atomic force microscopy (AFM) combined with a nanoneedle for targeted intracellular delivery.
Main Methods:
- Development of a nanoneedle system integrated with an atomic force microscope (AFM).
- Immobilization of DNA onto the nanoneedle surface using covalent bonding and avidin-biotin affinity.
- Confirmation of DNA immobilization through unbinding force measurements.
- In vitro testing of the DNA-nanoneedle system in HEK293 cells, including repeated insertion and withdrawal.
Main Results:
- Successful immobilization of DNA onto the nanoneedle was confirmed.
- The DNA-loaded nanoneedle was effectively inserted into HEK293 cells.
- Confocal microscopy and TO-PRO-3 iodide staining confirmed the presence of immobilized DNA on the nanoneedle.
- The immobilized DNA remained on the nanoneedle after multiple insertions and withdrawals from the cell.
Conclusions:
- The developed AFM-nanoneedle system offers a low-invasive method for single-cell gene and molecule transfer.
- The system demonstrates robust DNA immobilization and retention, even after repeated cellular manipulations.
- This technology holds promise for precise intracellular delivery and manipulation in biological studies.