RNA Transfection into Cultured Cells (In-Vitro) 

Scientists introduced the RNA transfection method, and for this method, they synthesize cationic lipid and then incorporate it or inject it into the liposome. They performed the transfection of Photinus Pyralis mRNA, an eastern firefly. This mRNA is luciferase, and they transfect into NIH 3T3 mouse cells cultured in-vitro. After this, cells showed luciferase activity. By observing the positive result of this experiment, this procedure can be implemented on humans, rats, mice, and other cultured cells. Scientists observed the spontaneous activity of beta-globin. By transfection, on NIH 3T3, it cleared that mRNA produces more luciferase protein due to untranslated beta globin than another mRNA lacking this component. This cationic liposome method is preferable to the DEAE dextran RNA transfection method. 

Transfection of Mononuclear Cells of Human Peripheral Blood with mRNA 

Peripheral blood of a human circulates in the whole body, and mononuclear cells comprise on single nucleus like monocytes. Scientists did in-vitro transfection of single nucleus cells of human peripheral blood with mRNA to stimulate the antigen-specific cytotoxic T lymphocytes to stimulate the cell-mediated immunity. The vaccines are needed to test to check the immune response and their efficacy. For this purpose, the target cells are necessary to check the vaccine. For the RNA-based or messenger RNA-based vaccines, the cells used to read the messenger RNA (mRNA) transfected monocytes which are mononuclear, derived dendritic cells. They transfect the mRNA into peripheral blood cells, and these cells are capable and start activating memory T cells in-vitro. Thus it proves that mRNA in-vitro transfection of peripheral blood mononuclear cells is an easy replacement of mRNA transfected monocyte dendritic cells in the laboratory or in-vitro to monitor or observe the immune response of the vaccines or to observe the natural immune response.