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Graphene oxide-based fluorescent "on/off" switch for visual bioassay using "molecular beacon"-hosted Hoechst dyes
Min Zhang1, Huynh-Nhu Le, Bang-Ce Ye
1Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology , Meilong Road 130, Shanghai 200237, China.
ACS Applied Materials & Interfaces
|August 24, 2013
Summary
A new molecular beacon (MB) probe enhances Hoechst dye fluorescence. This enables a graphene oxide (GO)-based "on/off" switch for sensitive, label-easy bioassays like DNA detection.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Materials Science
Background:
- Traditional molecular beacons (MBs) often require complex labeling and modification.
- Hoechst dyes are common fluorescent labels but can be difficult to control.
- Graphene oxide (GO) is known for its fluorescence quenching properties.
Discussion:
- A novel "molecular beacon"-hosted Hoechst dye (HMB) complex was designed, significantly enhancing Hoechst dye fluorescence.
- A graphene oxide (GO)-based fluorescent "on/off" switch was developed using HMB as signal indicators and GO as an energy acceptor.
- This system allows for efficient fluorescence quenching of HMB by GO in a label-easy format.
Key Insights:
- Demonstrated label-easy fluorescence "turn on" detection of sequence-specific DNA.
- Showcased label-easy fluorescence "turn off" detection of exonuclease activity.
- Achieved sensitive and selective detection in a single step within a homogeneous solution.
Outlook:
- The proposed design offers a cost-effective and simpler alternative to traditional MBs.
- Eliminates the need for fluorescence labeling or chemical modification of probes.
- Potential for broader applications in homogeneous bioassays and diagnostics.
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03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
02:11Reporter Genes
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
Commonly used reporter...