Telomere G quadruplex DNA (deoxyribonucleic acid) and thioflavin T mediated fluorescent biosensor and application thereof in lncRNA (long non-coding ribonucleic acid) detection

Abstract

The invention provides a telomere G-quadruplex DNA and thioflavin T mediated fluorescent biosensor and application thereof in lncRNA detection, and belongs to the technical field of fluorescence detection. The fluorescent biosensor at least comprises a capture probe, a signal probe, double-strand specific nuclease, terminal transferase and thioflavin T; wherein the capture probe is provided with a complementary region which is specifically hybridized with the lncRNA to be detected; the signal probe comprises a telomere G quadruplex DNA fragment and a poly T base chain. The fluorescent biosensor designed by the invention is used for sensitively detecting lncRNA in cells without marks on the basis of target recovery mediated by double-strand specific nuclease, amplification induced by terminal transferase and an enhanced fluorescence method induced by a thioflavin T / G-quadruplex compound, and has excellent sensitivity and specificity; and a valuable platform is provided for biomedical research, clinical diagnosis and treatment based on the lncRNA.

Description

technical field [0001] The invention belongs to the technical field of fluorescence detection, and in particular relates to a fluorescent biosensor mediated by telomeric G quadruplex DNA and thioflavin T and its application in lncRNA detection. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] Noncoding RNAs (noncoding RNAs, ncRNAs) are a class of RNA molecules that contain multiple types of RNA transcripts with protein-coding capabilities, and they participate in many biological processes, such as chromatin modification, post-transcriptional regulation, and epigenetics. ncRNAs include small non-coding RNAs (small non-coding RNAs, small ncRNAs) and long non-co...

AI Technical Summary

Problems solved by technology

Microarrays and RNA-sequencing facilitate novel RNA discovery, but they are limited by expensive instrumentation, poor specificity, and long analysis times

qRT-PCR and dPCR can sensitively detect low-abundance lncRNAs, but qRT-PCR requires precise control of reaction temperature through precision instruments, while dPCR involves complex sample pretreatment, expensive equipment and reagents

Recently, various isothermal amplification techniques have been introduced to detect miRNAs, including circle-mediated isothermal amplification, isothermal exponential amplification reaction, and rolling circle amplification, but they cannot directly detect lncRNAs because long lncRNAs cannot serve as primers

In fluorescence detection based on isothermal amplification, quenched Staudinger-triggered reporters, ssDNA reporters, and molecular beacons are often used for signal output, but the inventors found that they usually Limited by organic dyes and fluorescent labels, including incomplete self-quenching, poor stability, easy degradation, photobleaching, and high cost

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