g tetramer covalent coupling dna molecule and dna self-transfection kit and application

Abstract

The invention relates to a G-tetramer covalently coupled DNA molecule, the middle double strand is the double strand of the DNA fragment to be transfected, and the 5' end of each strand is G-connected by nine thymine (T) strands tetramer. The present invention also provides a DNA self-transfection kit, comprising polybrene, 5×Annealing buffer and the G tetramer covalently coupled DNA molecules. The invention is used for targeted DNA transfection of tumor cells, and selectively transfects genes or DNA fragments to be used into tumor cells. The transfection operation is simple, high efficiency and low toxicity, independent of cell cycle, simple operation and low cost.

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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