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A novel detection method for microRNA molecules in living cells

A detection method and technology for living cells, which are applied in biochemical equipment and methods, and the determination/inspection of microorganisms, which can solve the problems of low efficiency and insufficient sensitivity in the extraction process.

Active Publication Date: 2019-06-25
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a novel detection method for microRNA molecules in living cells, which solves the problems of low efficiency and insufficient sensitivity in the extraction process of current detection methods for microRNA molecules

Method used

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  • A novel detection method for microRNA molecules in living cells
  • A novel detection method for microRNA molecules in living cells
  • A novel detection method for microRNA molecules in living cells

Examples

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Embodiment 1

[0028] Example 1: A novel detection method for microRNA molecules in living cells

[0029] In the first step, 5 nm gold nanoparticles (GNP) modified with DNA1 and zinc cadmium selenium quantum dots (ZnCdSe) with an emission wavelength of 450 nm modified with DNA2-1 were programmed by DNA3-1 according to the rules of complementary base pairing assembled to obtain GNP-QD nano-assemblies; all DNA1 and DNA2-1 can carry out base pairing with DNA3-1;

[0030] In the second step, the GNP-QD assembly and DNA4-1 were transfected into cervical cancer cells (HeLa) using liposomes, and co-cultured for 6 hours;

[0031] In the third step, the HeLa cells were subjected to fluorescence imaging under a fluorescence microscope to detect whether they contained the target microRNA-21.

Embodiment 2

[0032] Example 2: A novel detection method for microRNA molecules in living cells

[0033] In the first step, 13 nm gold nanoparticles (GNP) modified with DNA1 and CdTe quantum dots (CdTe) with an emission wavelength of 550 nm modified with DNA2-2 are programmed according to the rules of complementary base pairing through DNA3-2. Assembled to obtain GNP-QD nano-assemblies, all DNA1 and DNA2-2 can perform base pairing with DNA3-2;

[0034] In the second step, the GNP-QD assembly and DNA4-2 were transfected into human breast cancer cells (MCF-7) using liposomes, and co-cultured for 6 hours;

[0035] In the third step, the MCF-7 cells were subjected to fluorescence imaging under a fluorescence microscope to detect whether they contained the target microRNA-203.

Embodiment 3

[0036] Example 3: A novel detection method for microRNA molecules in living cells

[0037] In the first step, the 30 nm gold nanoparticles (GNP) modified with DNA1 and the zinc-mercury-selenium quantum dots (ZnHgSe) with an emission wavelength of 750 nm modified with DNA2-3 were carried out according to the rule of complementary base pairing through DNA3-3. Procedurally assembled to obtain GNP-QD nano-assemblies; all DNA1 and DNA2-3 can perform base pairing with DNA3-3;

[0038] In the second step, the GNP-QD assembly and DNA4-3 were transfected into human breast cancer cells (MDA-MB-231) using liposomes, and co-cultured for 6 hours;

[0039] In the third step, the MDA-MB-231 cells were subjected to fluorescence imaging under a fluorescence microscope to detect whether they contained the target microRNA-141.

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Abstract

The invention relates to a novel method for detecting microRNA (microribonucleic acid) molecules in living cells. The method is characterized in that targeted microRNA molecules which are specifically expressed in cells are used as a catalyst to catalyze the disassembly reaction of the gold nanoparticle-quantum dot assembly, and the single microRNA molecule is repeatedly utilized to dissociate a plurality of QD and GNP, so that the previously quenched QD fluorescence signal is restored and subjected to multistage amplification, thereby implementing high-sensitivity detection and imaging on the targeted microRNA molecules in living cells, and providing references for distinguishing tumor cells from normal cells.

Description

technical field [0001] The present invention relates to the fields of analytical chemistry, nanomaterials, etc., and specifically relates to a novel detection method for microRNA molecules in living cells, which specifically includes microRNA molecules in living cells catalyzing the disassembly reaction between gold nanoparticles and quantum dots, changing GNP and The fluorescence resonance energy transfer effect between QDs can amplify the fluorescent signal of QDs, thereby realizing a highly specific and sensitive detection method for target microRNA molecules in living cells, and thus accurately distinguishing tumor cells from normal cells . Background technique [0002] Quantum dots (QDs for short) are semiconductor nanocrystals whose size is smaller than or similar to the exciton Bohr particle size. Compared with traditional organic molecular dyes, it has unique and excellent optical properties, such as wide excitation spectrum and continuous distribution, adjustable e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6818
CPCC12Q1/6818C12Q2525/207C12Q2563/137C12Q2563/155C12Q2565/101
Inventor 马楠何学文
Owner SUZHOU UNIV
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