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

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...

AI Technical Summary

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 produce a low signal. At the same time, the generation of molecular beacon fluorescence can only be linearly amplified, and the magnification is not high

Subsequently, the application of RT-PCR, rolling circle amplification, and constant temperature exponential amplification were all developed to detect miRNA. These methods have their own advantages, but complex label and probe design, and instrument requirements still limit their application.

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