MiRNA (Micro Ribonucleic Acid) detection probe and method for visually detecting miRNA

A technology for detecting probes and detection methods, which is applied in the fields of molecular biology and nucleic acid chemistry, and can solve problems such as low signal, limitation, and inability to fully open the stalk area, and achieve good response and good discrimination

Inactive Publication Date: 2012-08-01
WUHAN UNIV
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Problems solved by technology

However, the chip and various labeling methods all bring about the increase of detection cost
Subsequently, the application of molecular beacons brought a new approach to the detection of miRNA, but due to the limitation of the length of miRNA in the application of detecting miRNA, the length of the loop region of molecular beacon is limited to about 20-25 bases, so It is often impossible to fully open the stalk area and only produ...

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  • MiRNA (Micro Ribonucleic Acid) detection probe and method for visually detecting miRNA
  • MiRNA (Micro Ribonucleic Acid) detection probe and method for visually detecting miRNA
  • MiRNA (Micro Ribonucleic Acid) detection probe and method for visually detecting miRNA

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Embodiment 1

[0036] 1. Design of miRNA detection probe system

[0037] (1) The probe consists of three parts, G-quadruplex sequence, which is the signal part in the usual sense; interference sequence; and target complementary sequence, which is designed to specifically detect target miRNA. The probe is a hairpin structure under normal conditions, in which the interfering sequence is partially complementary to the 3′-terminal G-quadruplex forming sequence to form a double strand, and the complementary sequence of the miRNA to be tested is a loop loop.

[0038] (2) The detection target to be achieved: It can produce a good signal response to the corresponding target miRNA probe, the color is enough to be visible to the naked eye, and there is no background to the non-target miRNA. To achieve this purpose, a series of probes are designed, the chromogenic sequences of the probes are consistent, and the interfering sequences of the probes differ by one base.

[0039] 2. Verify the sensitivity ...

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Abstract

The invention relates to a miRNA (Micro Ribonucleic Acid) detection probe and a method for visually detecting miRNA. The miRNA detection probe comprises three parts: a G-quadruplex forming sequence close to a 3' end, a miRNA complementary sequence of a target to be measured in the middle, and an interference sequence at a 5' end. The probe has a hairpin structure under general conditions. After the target is added, the quantity of base pairs and stability of a DNA-RNA (Deoxyribonucleic Acid-Ribonucleic Acid) heterozygote are higher than that of a DNA double chain formed by the interference interface, and a hairpin structure is opened. The opened hairpin structure can release the G-quadruplex forming sequence under the action of a double-chain specific incision enzyme, G-quadruplex is formed under the action of sodium and potassium ions, and a catalyst with peroxidase activity is produced by assembling with Hemin. The method for detecting miRNA disclosed by the invention only requires one probe, a generated signal is visual, and expensive instruments are not required.

Description

technical field [0001] The invention belongs to the fields of molecular biology and nucleic acid chemistry, and relates to a miRNA detection probe and a method for visually detecting miRNA. Background technique [0002] MicroRNAs (miRNAs) are a class of non-coding RNAs with regulatory functions found in eukaryotic cells, usually about 20-25 nucleotides. miRNAs are formed by cleavage and processing of longer hairpin-containing precursors by Dicer enzymes, and then bind to RNA-induced silencing complexes (miRISCs) and recognize corresponding messenger RNAs (mRNAs) through complementary base pairing. Depending on the degree of complementarity, miRNAs can degrade or repress target mRNAs to repress the translation of target mRNAs. miRNA is widely expressed in animals and plants, and participates in various regulatory processes such as cell proliferation and apoptosis, development, virus defense, hematopoiesis, organ formation, and fat metabolism. miRNA is conserved, specific an...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 周翔王少儒田沺翁小成肖珩
Owner WUHAN UNIV
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