Optimization of transfection conditions for transfecting pPRMT1-shRNA into rat arterial endothelial cells

Objective To optimize the transfection conditions for transfecting the expression plasmids of short hairpin RNA(shRNA)targeted to the protein arginine methyltransferases 1(PRMT1)gene into primary cultured arterial endothelial cells mediated by polyethyleneimine jetPEITM-RGD. Methods The primary rat arterial endothelial cells was cultured using the method of attachment block,and identified with immunohistochemical SP method.Cells of the 3rd to 4th passages were chosen and transfected at the different proportions of jetPEITM-RGD and pPRMT1-shRNA.After 24 h,the transfection rate and cell viability were detected using fluorescent microscope and MTT assay,respectively. Results The transfection rate was highest(53.54%)when 3.0 μg pPRMT1-shRNA was added into each well of culture plate and the N/P ratio was equaled to 5. Conclusion This experiment may lay the foundation for performing efficient cell transfection and further research on RNA interfere of PRMT1 gene in vivo.