Cell Transfection Reagents

Transient transfection (transient transfection) is one of the ways to introduce a nucleic acid of interest into eukaryotic cells. In transient transfection, the nucleic acid of interest is introduced into a highly infectious cell line to obtain transient but high level expression of the nucleic acid of interest. The transfected nucleic acid of interest does not have to be integrated into the host chromosome, and the transfected cells can be harvested in a shorter time than stable transfection, and the expression of the gene of interest in the lysate can be detected.

At present, the products commonly used for transient transfection of cells on the market mainly include cationic liposomes or cationic polymer transfection reagents, however, such products have poor transfection effects on cells that are difficult to transfect, such as primary cells. In addition to the use of transfection reagents, the electroporation transfection technology that realizes transient transfection of cells through electroporation is also relatively common. Its transfection principle is to use instruments to form transient holes in the cell membrane when cells are stimulated by electric fields, so that exogenous molecules enter the cells to complete transfection. Although this method can realize the transfection of most cells including primary cells and other cells that are difficult to transfect, it does great harm to the cells, and the need to purchase special equipment, the operation steps are cumbersome and the transfection cost is high. Therefore, it is urgent to find a method for realizing transient transfection of cells, which has a wide application range, good transfection effect, no need to purchase special equipment, simple operation steps and low transfection cost.

 

this project found that under specific chemical treatment and pH treatment, microRNA can enter cells quickly and efficiently. On this basis, we further optimized the new transfection method and carried out in-depth research on the applicability of different cell lines, and developed a transfection reagent. The transfection reagent developed by the project has great application prospect in cell transfection (especially transient cell transfection), and has the following advantages:

• It can effectively transfect the target nucleic acid into different types of difficult-to-transfect cells such as HEK 293T cells, Hepa1 6 cells, primary hepatic cells and C2C12 cells, and has the advantage of a wide range of applications;
• When transfecting siRNA inhibiting the target gene in the host cell, the expression of the target gene can be significantly reduced within 6 to 12 hours of transfection, and the inhibition effect of the target gene is close to the effect of transfection using Lipofectamine 2000, which has the advantages of fast transfection onset time and good transfection effect;
• The whole transfection process of transfecting target nucleic acid in host cells does not need to use special equipment, the operation steps are simple, and the transfection cost is low;
• It has good stability and is conducive to market applications; the components used are all inexpensive, further reducing the cost of transfection.