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Transient transfection reagent and application method thereof

A technology of transient transfection and reagent, applied in the biological field, can solve the problems of strong anti-inflammatory reaction and toxicity, and achieve the effect of high transfection efficiency, low cost and high stability

Inactive Publication Date: 2019-05-10
WUHAN PROCELL LIFE SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, cationic liposomes have high efficiency in gene transfection in vitro, however, in vivo, it is rapidly cleared by serum, accumulates in lung tissue, and induces a strong anti-inflammatory response, which will lead to high levels of toxicity, so , which largely limits its application

Method used

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Examples

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Embodiment Construction

[0015] Below in conjunction with the examples, the specific implementation of the present invention will be further described in detail. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0016] According to the first aspect of the embodiments of the present invention, there is provided a transient transfection reagent comprising 2,4-pentanediol diacrylate and polyethyleneimine, and the pH of the solution is 7.05.

[0017] As a preference, dissolve branched PEI600 and 2,4-pentanediol diacrylate into 1.0 mL of anhydrous DMSO, mix thoroughly and react at room temperature for 6 h; stop the reaction with 0.5 mol / L HCl and adjust the pH to 4.5; place the polymer in HCl solution (pH4.0), and use a dialysis bag to dialyze overnight at 4°C to remove compounds with a relative molecular mass below 3000; freeze-dry the sample for later use;

[0018] Dilute the polymer and plasmid DNA with Opti MEM resp...

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Abstract

The invention provides a transient transfection reagent and an application method thereof. The reagent is prepared from 2,4-pentanediol dithiophosphate and polyethyleneimine, and pH of the solution is7.05. The transient transfection reagent is low in cost, high in transient transfection efficiency and stability, easy and convenient to operate, wide in application host cell range, and free of influence of antibiotic in a culture medium on the transfection efficiency.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a transient transfection reagent and an application method thereof. Background technique [0002] The cell membrane is a barrier that prevents extracellular substances from freely entering the cell, and it ensures the relative stability of the intracellular environment. It is precisely because the biological barrier function of the cell membrane prevents many biomacromolecular substances from entering the cell, which greatly limits the application of these substances in the field of therapy. Therefore, how to guide these substances to penetrate the cell membrane is an urgent problem to be solved. [0003] Gene transfection is "the transfer or transport of nucleic acid with biological function into cells, and the nucleic acid maintains its biological function in cells". There are many ways to introduce genes into eukaryotic cells, including biochemical transfection, physical tr...

Claims

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Application Information

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IPC IPC(8): C12N15/87
Inventor 冷毅斌吴亭
Owner WUHAN PROCELL LIFE SCI & TECH CO LTD