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Spherical nucleic acid probe and preparation method and application thereof

A nucleic acid probe, spherical technology, applied in the field of drug-loaded spherical nucleic acid probes and its preparation, can solve the problems of lack of targeting, lack of multifunctionality, etc., achieve good biocompatibility, high sensitivity, and simple preparation steps Effect

Inactive Publication Date: 2018-11-23
NANJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It lacks multifunctionality and lacks targeting in complex cellular bodies

Method used

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  • Spherical nucleic acid probe and preparation method and application thereof
  • Spherical nucleic acid probe and preparation method and application thereof
  • Spherical nucleic acid probe and preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0040] Human cervical cancer cells are used as detection and treatment objects, and then the spherical nucleic acid probe is used to detect telomerase activity and treat cancer cells.

[0041] Step 1, preparation of nano gold

[0042] The 13nm gold nano (AuNP) core was synthesized according to the reduction of chloroauric acid by sodium citrate. First, soak the glass instruments used in the experiment in aqua regia for 12 hours, and wash and dry them with a large amount of distilled water. Then, configure 100mL of 0.01% mass fraction of chloroauric acid solution, and transfer it to a 250mL three-necked flask and heat to boiling at 120°C. Then, 3.5 mL of 1% sodium citrate solution in mass fraction was quickly added under constant stirring. At the same time, keep boiling for 15 minutes and keep stirring. After the reaction, the reacted solution was cooled at room temperature and stirred continuously. Finally, store it in the refrigerator until later. figure 2 In order to p...

Embodiment 2

[0060] Human acute lymphoblastic leukemia T lymphocytes were used as the detection object and treatment object, and human acute lymphoblastic B cells were used as the control group. At the same time, TAMRA is used as the fluorescent signal group and BHQ-2 is used as the quenching group. Then, use the spherical nucleic acid probe to detect telomerase activity and treat cancer cells.

[0061] Prepare spherical nucleic acid probes according to steps 1 to 3 of Example 1. Because human acute lymphoblastic leukemia T lymphocytes are suspension cells, step 4 is different. First, cells in exponential growth phase are harvested by centrifugation. Then, its culture fluid is removed leaving a cell pellet. Then, the cells were washed twice with 0.01 M phosphate buffer (containing 0.1 M sodium chloride) and centrifuged. Finally, the cells were dispersed in 1 mL of CHAPS lysate (10 mM Tris-HCl, pH=7.4, 1 mM MgCl 2 , 1 mM EGTA, 1 mM PMSF, 10% glycerol, 0.5% CHAPS). Then, incubate at 4 °...

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Abstract

The invention discloses a spherical nucleic acid probe and a preparation method and application thereof. The spherical nucleic acid probe uses nanogold as a core of spherical nucleic acid; double-chain DNA (deoxyribonucleic acid) formed by hybridizing three chains and an adaptor are assembled to the surface of the nanogold to form an outer shell of the spherical nucleic acid probe. The spherical nucleic acid probe has the advantages that the activity of telomerase can be detected, and the DNA complex at the surface of the probe is rich in CG nucleotide and can carry an anti-drug medicine (adriamycin), so that the spherical nucleic acid probe can be used for treating the cancer cells, and can reach targeting property in the complicated biological environment by utilizing the modified adaptor at the surface of the probe; the stability is good, the biocompatibility is good, the toxic or side effect to the cells is avoided, the detection range is broad, the sensitivity is high, and the like; the detection speed is quick, and the preparation steps are simple.

Description

technical field [0001] The invention relates to the field of detection of telomerase activity and the field of drug loading, in particular to a drug-loading spherical nucleic acid probe and its preparation method and application. Background technique [0002] Telomerase is a special nucleoprotein reverse transcriptase. Grad and Blackburn first discovered telomerase in 1985. The telomerase holoenzyme is composed of a segment of RNA template (hTR), telomerase catalytic subunit (hTERT), dyskeratin, telomerase Cajal body protein 1 (TCAB1) and human reverse transcriptase protease. Telomerase can synthesize telomeric DNA repeat sequence (TTAGGG)n at the end of chromosome under the action of template RNA. Telomerase template RNA is composed of 451 nucleotides, and its template sequence is 3'-CAAUCCCAAUC-5'. Because telomerase prevents telomeres from shortening during cell division, telomerase plays a vital role in the infinite proliferation of cancer cells. In normal cells, tel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844C12Q1/48A61K47/54A61K47/69A61K31/704A61P35/00
CPCA61K47/549A61K47/6925A61P35/00A61K31/704C12Q1/48C12Q1/6844G01N2333/9128C12Q2521/113C12Q2525/151C12Q2563/107
Inventor 张卉杨国庆乔斌郭群群姜居倩
Owner NANJING NORMAL UNIVERSITY
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