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SOF nano gene vector for transfecting short-chain amino acids and preparation method and application of SOF nano gene vector

A gene carrier and short-chain nucleic acid technology, applied in the field of gene transfection, can solve the problems of high cytotoxicity, achieve mild synthesis conditions, convenient post-modification, and simple gene transfection methods

Inactive Publication Date: 2018-09-14
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to overcome the problem that the cationic liposome type gene carrier and the cationic polymer type gene carrier have relatively high cytotoxicity, and only have higher transfection efficiency to restricted cell lines and larger plasmid DNA, and provide a kind of gene carrier that can treat multiple Nano-gene carrier for high-efficiency gene transfection of small fragment nucleic acid in a kind of cell and its preparation method and application

Method used

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  • SOF nano gene vector for transfecting short-chain amino acids and preparation method and application of SOF nano gene vector
  • SOF nano gene vector for transfecting short-chain amino acids and preparation method and application of SOF nano gene vector
  • SOF nano gene vector for transfecting short-chain amino acids and preparation method and application of SOF nano gene vector

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Preparation of monomeric tetrahedral tetraphenylmethane derivatives.

[0046] The structure diagram of tetrahedral monomer molecule and CB[8] is shown in figure 1 , where D 1 Molecule as an example to illustrate the preparation process of tetrahedral tetraphenylmethane derivatives, Com-1~5 are contained in figure 1 in the described structure. The synthesis process can refer to the literature Nat. Commun. 2014, 5 , 5574. Tetrakis(4-bromomethylphenyl)methane (140 mg, 0.2 mmol) and isoquinoline (130 mg, 1 mmol) were weighed and dissolved in acetonitrile (30 mL), heated to reflux for 24 hours, and cooled naturally to At room temperature, thin-plate chromatography detected that the reaction did not change. Suction filter to obtain a solid, recrystallize with acetonitrile / methanol (5:1), filter and dry to obtain a light yellow solid, dissolve the solid in 20 mL of water, add saturated ammonium hexafluorophosphate solution (1 mL) to precipitate a solid, a...

Embodiment 2

[0047] Embodiment 2: Preparation of SOF nano gene carrier.

[0048] According to the report of our previous research work ( Nat. Commun. 2014, 5 , 5574 and Chinese Chem. Lett. 2017, 28 , 798), tetrahedral molecules with arylpyridinium salt groups at the end groups can form ordered 3D supramolecular organic frameworks (SOFs) in aqueous solution, and the stoichiometric ratio of tetraphenylmethane derivatives to CB[8] molecules Mix 1:2 in water, heat and ultrasonically disperse and dissolve the sample and cool to room temperature, then assemble in water to obtain the corresponding assembly, and then obtain the corresponding SOF nano-gene carrier.

Embodiment 3

[0049] Example 3: Characterization of the combination of SOF nanogene carrier and DNA with different base numbers in aqueous solution.

[0050] Taking the concentration of tetrahedral monomer molecules as the standard, prepare a SOF nanogene carrier solution with a solubility of 5 μM (tetrahedral molecules = 5 μM, CB[8] = 10 μM), keep SOF unchanged, add different proportions of DNA, and perform fluorescence tests. At the same time, the concentration of the SOF nanogene carrier was reduced by half (tetrahedral molecule = 2.5 μM, CB[8] = 5 μM), and different proportions of DNA were added for fluorescence testing. It can be confirmed by fluorescence test that the SOF nano-gene carrier has a good interaction with DNA. At the same time, according to its proportional relationship, it can be determined that its loading capacity can reach 150-400mg / g. After reducing the concentration, it still has a good loading capacity of 150-400mg / g. g. At the same time, a SOF nano-gene carrier so...

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Abstract

The invention belongs to the technical field of gene transfecting and particularly relates to an SOF nano gene vector for transfecting short-chain amino acids and a preparation method and applicationof the SOF nano gene vector. The SOF nano gene vector is SOF nano particles prepared by tetraphenylmethane serving as the core, the tetrahedron molecules, serving as the end groups, of aryl pyridiniumsalt groups or isoquinoline salt groups, and cucurbit[8]uril through self-assembling in water phase, the particle size of the particles is 10-300 nanometers, and the particles are adjustable in size;the SOF nano particles have mesoporous ducts serving as gene storage spaces. High interaction force exists between the positive charge porous feature of the SOF nano gene vector and nucleic acid molecules with negative charge, the SOF nano gene vector can effectively adsorb and combine with the nucleic acid molecules to form nano particles, the nano particles can be phagocytized by cells, and treatment genes can diffuse and transfer into cell nucleuses after leaving frame structures. The positive ion pores of the SOF nano gene vector have a quite good transfecting effect on nucleic acid of certain length in various cells.

Description

technical field [0001] The invention belongs to the technical field of gene transfection, and in particular relates to a nano-gene carrier capable of effectively transfecting various cells with genes, a preparation method and application thereof. Background technique [0002] Gene therapy is the introduction of therapeutic genes or targeted genes into target cells, so that the therapeutic genes or targeted genes are expressed in the cells, thereby treating diseases caused by gene abnormalities or defects. It is a new treatment in the field of medical treatment research. It has been widely used in various diseases such as cancer, genetic diseases, cardiovascular diseases, infectious diseases and autoimmune diseases. Gene therapy is divided into three parts: the selection of therapeutic genes or targeted genes, the introduction of therapeutic genes or targeted genes into target cells, and the expression of therapeutic genes or targeted genes in target cells. The most critical...

Claims

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Application Information

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IPC IPC(8): C12N15/87
CPCC12N15/87
Inventor 黎占亭周璐杨波张晓丹张丹维王辉刘传志祁琦闫萌赵亚坤
Owner FUDAN UNIV
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