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A nucleic acid nanostructure and its preparation method and application

A nanostructure and nucleic acid nanotechnology, applied in the field of DNA nanotechnology, can solve problems such as weak electromagnetic field strength, DNA origami structure size, function and complexity limitations, poor stability of precious metal nanostructures, etc., to achieve large size, diverse shapes, and stability Good results

Active Publication Date: 2020-12-15
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method also has obvious defects. Due to the influence of the single-strand length of the circular phage and the fixed base sequence, the size, function and complexity of the DNA origami structure are obviously limited, and the stability of the assembled noble metal nanostructure is relatively low. Poor, the generated electromagnetic field strength is weak

Method used

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  • A nucleic acid nanostructure and its preparation method and application
  • A nucleic acid nanostructure and its preparation method and application
  • A nucleic acid nanostructure and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0078] The modification of embodiment 2 golden triangular prism

[0079] Treat the thiolated oligonucleotide and the thiolated PEG-400 with a concentration of 200 mM TCEP for six hours, respectively, to reduce the disulfide bonds in the oligonucleotide and PEG-400 to monothiol; use a size exclusion column ( G-25, GE Healthcare) to purify oligonucleotides and remove TCEP; the purified DNA and PEG were added in a molar ratio of 10000:1 and 1000:1 to the OD value containing 0.01% (w / v) SDS 1 in the gold nanoprism solution; after incubating the mixed solution at 30°C for 6 hours, slowly add a NaCl solution with a concentration of 5M so that the final concentration of NaCl is 500mM; Centrifuge for 10 minutes, add 1 mL of 0.5× TBE buffer containing 200 mM NaCl to resuspend, and repeat the centrifugation three times to remove excess thiolated DNA.

Embodiment 3

[0080] Synthesis of Example 3 Nucleic Acid Nanostructure

[0081] First, prepare six conventional origami structures:

[0082] Mix the M13 backbone and corresponding staple strands, capture strands, and linker strands at a ratio of 1:10, and anneal using a gradient PCR machine. The reaction system was: 75 μL ultrapure water, 10 μL 10×TAE / Mg buffer solution, 5 μL M13 main chain, 10 μL mixture of staple strand and capture strand; the annealing procedure was: (1) from 95°C to 70°C, ℃ is a gradient, and each gradient stays for 5 minutes; (2) From 70°C to 50°C, every 2°C is a gradient, and each temperature gradient is kept for 10 minutes; (3) From 50°C to 20°C, every 1°C For one gradient, each gradient stays for 15 minutes, and the entire annealing process takes about 10 hours in total.

[0083] After performing the set annealing program, the triangular origami structure was centrifuged with a 100kDa spin column (MWCO) to remove excess capture strands and staple strands. The spe...

Embodiment 4

[0085] Example 4 Assembly and purification of nucleic acid nanostructures and gold triangular prisms

[0086] Mix hexagonal nucleic acid nanostructures with DNA-functionalized gold triangular prisms at a molar ratio of 1:20 for annealing assembly: from 45°C to 25°C, every 1°C is a gradient, each gradient stays for 7 minutes, cycle 8 times, for about 24 hours, to get a golden bow structure.

[0087] The annealed product was electrophoresed on an agarose gel stained with 1.0% ethidium bromide in a running buffer containing 12 mM MgCl 2 1×TBE, the loading buffer was 50% glycerol, the voltage was 15V / cm, and the electrophoresis time was 1 hour. The band of interest was cut and the golden bow-tie structure was extracted from the gel with a Freeze-Squeeze column (Bio-Rad) at 4°C.

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Abstract

The invention provides a nucleic acid nanostructure and its preparation method and application. The nucleic acid nanostructure is a hexagonal DNA nanostructure formed by assembling six triangular DNA origami structures constructed by DNA origami technology, specifically through scaffolding chains and The staple strand and the capture strand are hybridized, and the hexagonal DNA nanostructure is self-assembled by hybridizing the connecting strands with the scaffold strands of the six triangular DNA origami structures. The nucleic acid nanostructure has good stability and can anchor two 80nm gold triangular prisms, and the prepared gold bowtie structure can generate a strong electromagnetic field, realizing single-molecule Raman detection.

Description

technical field [0001] The invention belongs to the field of DNA nanotechnology, and relates to a nucleic acid nanostructure and its preparation method and application. Background technique [0002] Under the action of light, electrons on the surface of noble metal nanostructures absorb photon energy, and localized surface plasmon resonance (LSPR) occurs, generating a strong electromagnetic field. The wavelength and field strength at the peak of the LSPR spectrum depend on the shape, size, geometric arrangement, and physicochemical properties of the noble metal nanostructures. Studies have shown that strong electromagnetic fields can be generated between noble metal nanomaterials with reasonable structures. This property has great application potential in the field of optics. It has been applied in optical tweezers, single-molecule fluorescence detection, surface-enhanced Raman scattering and nonlinear optics. etc. [0003] Among the many metal nanostructures, the gold bow...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10B82Y5/00B82Y40/00G01N21/65B22F9/24B22F1/00
CPCG01N21/65C12N15/10C12N15/11B82Y5/00B82Y40/00B22F9/24G01N2021/656B22F1/0553B22F1/054C12Q1/68
Inventor 丁宝全湛鹏飞李娜王振刚蒋乔王婷
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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