G-tetramer covalently coupled DNA molecule, DNA self-transfection kit and application

Abstract

The present invention relates to a G-tetramer covalently coupled DNA molecule. The middle double strands are double strands of a to-be-transfected DNA fragment, and the 5' end of each strand is G-tetramer formed by strain connection of 9 T (thymine). The invention further provides a DNA self-transfection kit. The kit comprises polybrene, 5*Annealing buffer and the G-tetramer covalently coupled DNAmolecule. The G-tetramer covalently coupled DNA molecule and the DNA self-transfection kit are used for targeted DNA transfection of tumor cells, and genes or DNA fragments with specific applicationare transfected into tumor cells. The G-tetramer covalently coupled DNA molecule and the DNA self-transfection kit have the advantages of being efficient, low in toxicity, independent of cell cycle, simple to operate and low in cost, and transfection operation is simple and convenient.

Description

technical field [0001] The invention relates to a DNA transfection technology, in particular to a G tetramer covalent coupling DNA molecule and a DNA autotransfection kit and application. Background technique [0002] Transfection refers to the process of introducing exogenous nucleic acid into cells. There are three main types of transfection methods at present, namely physical, chemical and biological methods. Physical methods mainly refer to electroporation and microinjection, which require equipment to achieve. Transfection, and the cell survival rate of the electroporation method is low, the operation of the microinjection method is complicated, and the equipment is expensive. The chemical approach mainly includes cationic liposomes, cationic polymers and calcium phosphate and other types of transfection reagents, which all need to rely on the endocytosis of cells. Although the cationic liposome method has a wide range of applications, it has certain cytotoxicity and is...

AI Technical Summary

Problems solved by technology

[0002] Transfection refers to the process of introducing exogenous nucleic acid into cells. There are three main types of transfection methods at present, namely physical, chemical and biological methods. Physical methods mainly refer to electroporation and microinjection, which require equipment to achieve Transfection, and the electroporation method has a low cell survival rate, the microinjection method is complicated to operate, and the equipment is expensive

The chemical approach mainly includes cationic liposomes, cationic polymers and calcium phosphate and other types of transfection reagents, which all need to rely on the endocytosis of cells. Although the cationic liposome method has a wide range of applications, it has certain cytotoxicity and is harmful to cells. The concentration of DNA is required, too high a DNA concentration will affect the transfection efficiency, the repeatability of the calcium phosphate method is poor, and the concentration of DNA is high, and reagents including cationic polymers are dependent on cell mitosis

Biological approaches mainly include retroviral methods, which, although highly efficient, are immunogenic and only infect dividing cells

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