Low Effectiveness of the Introduction of pmaxGFP into Primary Human Coronary Endothelial Cells Using Cell-Penetrating Peptides and Nuclear-Localization Sequences in Non-Covalent Interactions

Abstract

Cell-penetrating peptides (CPPs), due to their effectiveness and low cytotoxicity, are of increasing interest in the context of the transport of macromolecules to the cells. The simplest and safest method seems to be the non-covalent binding of CPP and cargo molecules. However, it requires the optimization of the reaction conditions. The study aimed to determine the effectiveness and cytotoxicity of the Pep-1, KALA, and TAT proteins as well as the NLS [47–55] and NLS [47–56] sequences derived from the Simian Vacuolating 40 (SV40) T-antigen in the context of the transport of the pmaxGFP plasmid to primary human coronary artery endothelial cells. The results are presented in the form of extensive photographic documentation, which shows significant differences in the efficiency of the transfection process between electroporation and the use of CPPs. The study presents negative results in which, despite the manipulation of various parameters (incubation time, incubation temperature, culture time, charge ratio, plasmid concentration), results similar to electroporation were not obtained.

Keywords

• cell-penetrating peptides
• primary human coronary artery endothelial cells
• TAT
• KALA
• Pep-1