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CN-121975809-A - Palindromic mediated self-assembled DNA molecule, and preparation method and application thereof

CN121975809ACN 121975809 ACN121975809 ACN 121975809ACN-121975809-A

Abstract

The invention discloses a palindromic mediated self-assembled DNA molecule, a preparation method and application thereof, and belongs to the technical field of biology. The palindromic mediated self-assembled DNA molecule is obtained by self-assembling an A1 chain, an L1 chain and a C1 chain, wherein the nucleotide sequence of the A1 chain is shown as SEQ ID NO.1, the nucleotide sequence of the L1 chain is shown as SEQ ID NO.2, and the nucleotide sequence of the C1 chain is shown as SEQ ID NO. 3. The palindromic mediated self-assembled DNA molecule A1-L1-C1 can be used for detecting miR-21, and has higher detection sensitivity and specificity. The A1-L1-C1 also has a drug carrying function, can target specific tumor cells, and can accurately deliver drugs into cancer cells. The invention provides a new technical approach for early diagnosis of tumor, detection of intracellular biomolecules and targeted drug delivery, and has wide application prospect.

Inventors

  • LI MIN
  • Wu Denghang
  • PAN WENHAO
  • LI WEI

Assignees

  • 温州医科大学

Dates

Publication Date
20260505
Application Date
20260209

Claims (10)

  1. 1. A palindromic self-assembled DNA molecule, wherein said palindromic self-assembled DNA molecule is self-assembled from A1 chain, L1 chain and C1 chain; The nucleotide sequence of the A1 chain is shown as SEQ ID NO.1, and the 5' end of the A1 chain is modified with a fluorescent group; the nucleotide sequence of the L1 chain is shown as SEQ ID NO.2, and the 3' end of the L1 chain is modified with a quenching group; The nucleotide sequence of the C1 chain is shown as SEQ ID NO.3, and a fluorescent group is modified between the 10 th base and the 11 th base.
  2. 2. The method for preparing the palindromic self-assembled DNA molecule according to claim 1, characterized in that the palindromic self-assembled DNA molecule is obtained by mixing the A1 chain, the L1 chain and the C1 chain at a molar ratio of 1:1:0.7 and then reacting at 90-98℃for 4-6 min.
  3. 3. The use of the palindromic self-assembled DNA molecule of claim 1 for the preparation of a product for detecting miR-21, wherein said product comprises a reagent, a kit or a chip.
  4. 4. A product for detecting miR-21, comprising the palindromic self-assembled DNA molecule of claim 1.
  5. 5. Use of the palindromic self-assembled DNA molecule according to claim 1 for the preparation of a tumor-targeted drug delivery system.
  6. 6. A method for preparing a tumor targeting drug delivery system, which is characterized in that the palindromic self-assembled DNA molecule of claim 1 is mixed with a drug for incubation, thereby obtaining the tumor targeting drug delivery system.
  7. 7. The method according to claim 6, wherein the incubation is performed at a temperature of 20-25 ℃ for a time of 10-14h.
  8. 8. A tumor-targeted drug delivery system prepared by the preparation method of claim 6 or 7.
  9. 9. Use of the tumor-targeted drug delivery system of claim 8 in the preparation of a medicament for treating tumors.
  10. 10. Use of a palindromic mediated self-assembled DNA molecule according to claim 1 for the preparation of a product for tumor imaging.

Description

Palindromic mediated self-assembled DNA molecule, and preparation method and application thereof Technical Field The invention relates to the technical field of biology, in particular to a palindromic mediated self-assembled DNA molecule, a preparation method and application thereof. Background According to global cancer data issued by the world health organization (World Health Organization, WHO) in 2020, the number of cases of malignant tumors and the number of cases of death are increased year by year, and become one of the main causes of death for citizens in all countries of the world. Therefore, there is an urgent need to control the occurrence and development of cancer, thereby effectively reducing the morbidity and mortality. To achieve optimal therapeutic effect, it is necessary to discover the tumor early. At present, research related to early detection of tumors has become a hot spot problem in the medical field. Nucleic acids have been widely used as an excellent structural material for constructing various types of nucleic acid nanomaterial molecules. The nucleic acid nano material is very suitable for the biomedical field due to the characteristics of excellent biocompatibility, biodegradability, safety, convenient automatic synthesis, good programmability and the like. DNA is subjected to A-T, G-C Watson Crick base pairing, so that the DNA has the characteristics of programmability and addressability, and therefore, in order to prepare nucleic acid nano structures with various functions, the DNA nano structures can be prepared by designing and utilizing the rigidity of double strands and the flexibility of single strands of the DNA. In addition, DNA is an endogenous molecule of an organism, and the constructed functional nucleic acid nanomaterial of the DNA shows good compatibility in the external environment and biological system in vivo. After the recent 40 years of development of nucleic acid nanotechnology, scientists have been able to assemble many complex and fine two-dimensional, three-dimensional, and even curved specific self-assembled structures. Nucleic acid nanostructures formed by self-assembly are easily modified, and many nucleic acid nanostructures can modify specific nucleic acid aptamers to perform specific functions. Microribonucleic acid (miRNA) is taken as one of exosomes and is utilized by tumor cells to promote the growth and metastasis of the tumor cells, so that the high death rate of tumor patients is finally caused. Therefore, miRNA is used as an index for early diagnosis of tumors, and the change of miRNA content in the occurrence and development processes of tumor cells is detected, so that the method has very broad clinical application prospect. microRNA-21 (miRNA-21/miR-21) has been shown to be aberrantly expressed in almost all tumors, such as lung adenocarcinoma, B-cell lymphoma, breast cancer, myelogenous leukemia, lung cancer, cholangiocarcinoma and other cancers, and to promote metastasis. Current literature indicates that miR-21, as an oncogenic miRNA, participates in cell growth, metastasis and apoptosis by controlling signaling pathways and various target molecules, and can produce negative regulatory effects on target genes by promoting mRNA degradation of the target genes or inhibiting mRNA translation. Notably, increasing evidence further demonstrates that miR-21 is closely related to diagnosis, recurrence and prognosis of cancer patients, suggesting that miR-21 may be a novel biomarker for diagnosis and prognosis of tumors. However, the existing detection means for miR-21 have the defects of low sensitivity and poor specificity, and the clinical application of miR-21 serving as a biomarker is limited. Disclosure of Invention The invention aims to provide a palindromic mediated self-assembled DNA molecule, a preparation method and application thereof, so as to solve the problems in the prior art. The invention constructs a palindromic mediated self-assembled DNA molecule A1-L1-C1, wherein the A1-L1-C1 can be used for detecting miR-21, and has higher detection sensitivity and specificity. The A1-L1-C1 also has a drug carrying function, can target specific tumor cells, and can accurately deliver drugs into cancer cells. The invention provides a new technical approach for early diagnosis of tumor, detection of intracellular biomolecules and targeted drug delivery, and has wide application prospect. In order to achieve the above object, the present invention provides the following solutions: The invention provides a palindromic mediated self-assembled DNA molecule, which is self-assembled by an A1 chain, an L1 chain and a C1 chain; The nucleotide sequence of the A1 chain is shown as SEQ ID NO.1, and the 5' end of the A1 chain is modified with a fluorescent group; the nucleotide sequence of the L1 chain is shown as SEQ ID NO.2, and the 3' end of the L1 chain is modified with a quenching group; The nucleotide sequence of the C1 chain is shown as SEQ ID NO.3