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Biosensor for targeting cancer cells based on DNA triangular prism structure conformation change

A technology for biosensors and cancer cells, applied in the field of biosensors, can solve the problems of excessive nanostructure stability and low space utilization, and achieve the effects of fast detection speed, fast response speed, and sensitive detection.

Active Publication Date: 2021-02-09
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the development of DNA nanotechnology, many DNA nanoassemblies such as DNA origami, DNA tetrahedron, and DNA cage have been successfully constructed and applied to targeted imaging of cancer cells, but these nanostructures are too stable and have low space utilization

Method used

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  • Biosensor for targeting cancer cells based on DNA triangular prism structure conformation change
  • Biosensor for targeting cancer cells based on DNA triangular prism structure conformation change
  • Biosensor for targeting cancer cells based on DNA triangular prism structure conformation change

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of triangular prism nanostructure

[0036] (1) Synthesize T1, T2, S1, S2, S3, and S4 according to the sequence of SEQ ID NO: 1-6, wherein the 5' and 3' ends of S4 are respectively modified with CY3 and CY5;

[0037] With 1× TAE solution (pH 8.0, containing 10 mM Mg 2+ ) were prepared into dilutions with a concentration of 100 µM;

[0038] (2) Take sterilized EP tubes, add 5 μL of T1, T2, S1, S2, S3, and S4 solutions respectively, make up to 50 μL with sterilized water, keep warm at 95°C for 10 min, gradually cool to room temperature, and anneal to obtain three Solution (10 µM) of prismatic nanostructures.

Embodiment 2

[0039] Embodiment 2 detects the screening of pH condition

[0040] (1) Configuration buffer, containing MgAc 2 (10 mM), KAc 2 (140 nM), EDTA (0.1 mM), the pH of the solution was adjusted to 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5 and 8 with acetic acid or potassium hydroxide, respectively.

[0041] (2) Add 5 µL of buffer solutions with different pHs to 9 sterilized EP tubes, add 5 µL of the solution containing triangular prism nanostructures in Example 1, and shake for 30 s;

[0042] (3) Add 5 μL of 10 mM ATP solution, and place the centrifuge tube in a water bath at 37°C for 2 h; after the reaction, use F-4600 fluorescence spectrum detection at room temperature (excitation wavelength 525 nm, fluorescence emission spectrum scanning range 550 -750 nm, slit 1 nm);

[0043] The fluorescence intensity at 667nm of the pH 4.5 buffer is 1, and the normalized fluorescence intensity is calculated. See the results in figure 2 , it can be seen from the figure that the detected fluorescent si...

Embodiment 3

[0044] Example 3 ATP concentration screening

[0045] (1) Add 5 µL of the pH 5 buffer solution prepared in Example 2 to 12 sterilized EP tubes, add 5 µL of the solution containing triangular prism nanostructures in Example 1, and shake for 30 s;

[0046] (2) Add 5 μL of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1, 2, 4, 6, 8, 10 mM ATP solution, and place the centrifuge tube in a water bath at 37°C for 2 h; At room temperature, use F-4600 fluorescence spectrum detection (excitation wavelength 525 nm, fluorescence emission spectrum scanning range 550-750 nm, slit 1 nm);

[0047] Taking the fluorescence intensity at 667 nm of the test solution with an ATP concentration of 10 mM as 1, the normalized fluorescence intensity was calculated, and the results are shown in image 3 , it can be seen from the figure that the detected fluorescence signal increases as the concentration of ATP increases in the range of 0-10 mM, and when the concentration reaches 10 mM, the fluorescence signal reaches the...

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Abstract

The invention belongs to the technical field of biosensors, and relates to a biosensor for targeting cancer cells based on DNA triangular prism structure conformation change. Before detection, a DNA triangular prism nanostructure is firstly constructed; then the pH value of a reaction solution is changed to change the conformation of the triangular prism nanostructure; and a cancer marker ATP is added, proximity reaction is performed to enable two fluorophores to be close to generate FRET, and detection is performed through a fluorescence spectrum. The sensor has the advantages of high detection speed, low detection limit, high specificity and the like, can make up for the defects and deficiencies of the existing detection method, and realizes rapid and accurate quantitative detection.

Description

technical field [0001] The invention belongs to the technical field of biosensors, and relates to a biosensor targeting cancer cells based on conformational changes of DNA triangular prism structures. Background technique [0002] An excellent nanostructure should have good imaging and drug delivery capabilities at the same time in order to maximize its application value in tumor diagnosis and treatment. Compared with exogenous materials, DNA has good biological affinity and is the main material for biotherapeutic and biomedical applications. With the development of DNA nanotechnology, many DNA nanoassemblies such as DNA origami, DNA tetrahedron, and DNA cage have been successfully constructed and applied to targeted imaging of cancer cells, but these nanostructures are too stable and have low space utilization. Therefore, it is urgent to develop a DNA nanostructure with strong specificity and controllable structure for target cell imaging. Contents of the invention [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6456G01N21/64G01N2021/6439G01N2021/6417
Inventor 黄加栋王业茹王玉刘素李明涵孙文玉张曼茹江龙
Owner UNIV OF JINAN
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