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Preparation method and application of two-photon deoxyribozyme metal organic framework probe

A metal-organic framework and deoxyribozyme technology, applied in the field of nanomaterial manufacturing, can solve the problems of zinc ion deoxyribozyme imaging effect and penetration depth, high cell entry efficiency, poor biological stability, etc., and achieve high selectivity Two-photon fluorescence imaging, strong membrane penetration ability, and high loading effect

Active Publication Date: 2020-01-10
ZHENGZHOU UNIV
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Problems solved by technology

[0006] The present invention proposes a two-photon deoxyribozyme metal-organic framework to solve the problems of unsatisfactory imaging effect and penetration depth of the existing zinc ion deoxyribozyme, weak membrane penetration ability, poor biological stability and difficulty in being used in complex biological systems. The preparation method and application of the probe can be applied to imaging of living cells and apoptotic cells. This application is simple to operate, has little light damage to the sample, low background interference, high sensitivity, and high cell entry efficiency

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  • Preparation method and application of two-photon deoxyribozyme metal organic framework probe
  • Preparation method and application of two-photon deoxyribozyme metal organic framework probe

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Embodiment 1

[0048] A method for preparing two-photon deoxyribozyme metal-organic framework nanoprobe (TP-Zn-ES-MOF), the preparation principle is as follows figure 1 Shown: in Zr 4+ and H 2 BDC is used as the raw material to synthesize MOF with regular octahedral structure, and then loaded with zinc ion deoxyribozyme double-stranded TP-Zn-ES, in which the substrate chain is labeled with two-photon fluorophore TP, and the enzyme chain is labeled with a quencher group (BHQ1). The zinc ion probe TP-Zn-ES-MOF was obtained. The experimental steps are as follows:

[0049] The two-photon fluorophore TP was synthesized by DNA solid-phase synthesis (the synthetic route is shown in the appendix figure 2 ) was labeled to the 5-end of the zinc ion deoxyribozyme substrate chain (Zn-S) to obtain TP-Zn-S; Enzyme chain (BHQ1-Zn-E) (molar ratio 1:2) was mixed into Tris buffer solution (25 mM Tris, 50 mM MgCl 2 , 100 mM NaCl, pH 7.4), heated to 95°C for 10 minutes, and then annealed and cooled on ice...

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Abstract

The invention relates to a preparation method and application of a two-photon deoxyribozyme metal organic framework probe. The method comprises the following steps: marking a two-photon fluorophore ona substrate chain of zinc ion deoxyribozyme, marking a quenching group on an enzyme chain to obtain a two-photon deoxyribozyme probe TP-Zn-ES, and synthesizing a carrier-metal organic framework (Zr-MOF) by using zirconium tetrachloride and terephthalic acid as raw materials through a solvothermal method; and supporting the two-photon deoxyribozyme probe TP-Zn-ES on the the Zr-MOF carrier to obtain the TP-Zn-ES-MOF probe. The method is simple to operate, green and environmentally friendly, and creats a novel strategy for preparing the two-photon deoxyribozyme metal organic framework probe; andthe TP-Zn-ES-MOF probe prepared by taking zinc ions as examples can be used for imaging of zinc ions in living cells and apoptotic cells, and shows huge application value in the fields of medical diagnosis and treatment.

Description

technical field [0001] The invention belongs to the field of nanomaterial manufacturing, and relates to the preparation and application of a DNAzyme specific recognition ability, two-photon fluorescence imaging technology and a metal organic framework carrier, in particular to a two-photon DNAzyme metal organic framework probe The preparation method and its application. Background technique [0002] Deoxyribozyme is a kind of catalytic nucleic acid with specific recognition function obtained through in vitro screening. It has many advantages such as biocompatibility, water solubility, low toxicity, and easy synthesis and functionalization. Among them, RNA-cleaving DNase, as the most widely used DNase, is of great significance in biological analysis and biomedical technology. (Juewen Liu, ZehuiCao, and Yi Lu, Chem. Rev.2009, 109, 5, 1948-1998) However, RNA-cleaving DNAzymes have defects in cell applications: on the one hand, due to the negatively charged DNAzymes and the cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/44G01N21/64
CPCC12Q1/44G01N21/6486
Inventor 孟红敏石鑫鑫吴亚楠葛佳李朝辉
Owner ZHENGZHOU UNIV
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