DNA zipper molecule modified lipidosome vesicle and preparation method and application thereof

Abstract

The invention provides a DNA zipper molecule modified lipidosome vesicle and a preparation method and application thereof. The lipidosome vesicle is composed of a DNA zipper molecule modified bimolecular layer membrane and a molecular probe wrapped in the bimolecular layer membrane, wherein DNA zipper molecules have a DNA zipper sequence promoting lipidosome fusion, the lipidosome vesicle is rapidly subjected to membrane fusion with an exosome modified by the DNA zipper molecules under the action of a DNA zipper, so that the molecular probe contained in the vesicle is sent into the exosome, and the in-situ, accurate and rapid detection of a nucleic acid-containing biomarker in the exosome is realized. The preparation method of the liposome vesicle comprises the steps: mixing the bimolecular layer membrane and the molecular probe, extruding to obtain the liposome vesicle, and incubating the liposome vesicle with the DNA zipper molecule modified with a lipophilic group. The DNA zipper molecule modified on the surface of the liposome vesicle is actively fused with the exosome, so that the efficiency is higher, the nucleic acid marker to be detected does not need to be separated from the exosome in advance, and the detection effect is better.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a liposome vesicle modified by DNA zipper molecules, a preparation method and application thereof. Background technique [0002] Exosomes have received more and more attention because they contain important intercellular information exchange molecules in organisms and can be used in many fields such as biological detection, diagnosis and treatment. Exosomes originally refer to 40-100nm vesicle-like structures secreted during reticulocyte differentiation and formed by the fusion of multivesicular bodies and plasma membranes. The outer layer of exosomes is a phospholipid bilayer, which contains proteins, mRNA, miRNA, DNA fragments, etc. related to the source cells, which is an important basis for exosomes to participate in the exchange of information and substances between cells. Exosomes are widely present in blood, saliva, urine, tears, and cerebrospinal fluid, e...

AI Technical Summary

Problems solved by technology

However, the transmembrane efficiency of simple molecular beacons is low, and the accuracy of detection may not be guaranteed

The diffusion efficiency of molecular beacons can be improved by adding enzymes that increase the permeability of exosome membranes, but at the same time it will cause the leakage of biomarkers in exosomes, which will bring errors to the detection

At the same time, there may be free miRNA in the extracted exosome solution, and direct co-cultivation of hairpin-structured molecular beacons with exosomes may interfere with the determination of the actual miRNA content in exosomes, and the exposed molecular beacons may be absorbed by body fluids. Nucleic acid degradation, affecting the accuracy of detection

Based on the above analysis, the existing techniques for detecting nucleic acid molecules in exosomes using molecular probes have the following problems: (1) Exosomes need to be isolated in advance, the isolation steps are lengthy, and there are various types of extracellular vesicles, and the isolated products often contain other types vesicles and protein aggregates, the separation step will also cause damage to exosomes; (2) the efficiency of molecular probes entering exosomes through diffusion is low, and they are easily decomposed by nucleases in body fluids, which may generate non-specific signals , so it is difficult to achieve accurate and efficient detection of nucleic acid molecules in exosomes

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