Ratiometric viscous flare fluorescent probe as well as preparation method and application thereof

Abstract

The invention discloses a ratiometric viscous flare fluorescent probe as well as a preparation method and application thereof. The probe uses gold nanoparticles as a carrier, and the surface of the carrier is connected with DNA (Deoxyribonucleic Acid) double chains with different fluorescent groups for identifying telomerase: FL-R flare long chain and SH-C-FL strand short chain with a partial complementary sequence of the long chain. The FL-R flare long chain comprises fluorescent dye, an auxiliary sequence, a complementary sequence of a telomere amplification sequence and a telomere primer sequence in sequence from a 5'end. No matter the telomere exists or not, the FL-R flare long chain is luminescent, and can be used as an internal fluorescence signal reference. The SH-C-FL strand shortchain is modified with sulfydryl at one end and modified with another fluorescent dye at the other end. In the presence of the telomere, the short chain is prolonged to form a hairpin structure and falls off from nanogold, and fluorescence is opened for in-situ detection of telomere activity in cells. In combination with a long-chain fluorescence signal, quantitative analysis of telomere activityin cancer cells and effective differentiation of tumor cells from normal cells can be realized. The probe can be directly applied to laser confocal imaging without cleaning during cell experiments.

Description

technical field [0001] The invention relates to the field of analysis detection and medical diagnosis, in particular to a ratio-type viscous flare fluorescent probe for in-situ detection of telomerase activity in cells, a preparation method and application thereof. Background technique [0002] Telomeres are special structures located at the ends of chromosomes that prevent chromosome degradation, rearrangement, and end fusion, thereby maintaining the integrity of the genome. Studies in recent years have found that the continuous shortening of telomere length during cell division is the cause of "replicative aging" of somatic cells, and in tumor cells, overactivated telomerase can repair and prolong the shortened telomere , so that telomeres will not be lost due to cell division, so that the number of cell divisions will increase. Telomerase is inactive in normal somatic cells, but is overactivated in almost all tumor cells, maintaining the immortal phenotype or infinite pr...

AI Technical Summary

Problems solved by technology

[0004] In order to solve the technical problems existing in the existing telomerase detection means, the present invention designs and synthesizes a series of ratiometric viscous flares with low fluorescence background, high specificity, good membrane penetration and quantitative detection of telomerase activity (sticky-flare) fluorescent probe and its preparation method and application

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