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

Wioletta Zielińska,Maciej Gagat,Klaudia Mikołajczyk,Marta Hałas-Wiśniewska,Alina Grzanka

doi:10.3390/app11051997

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.

Highlights

IntroductionPublisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations
Pictures included in the publication show representative views for all of the experiments
For greater readability, the results are presented in the form of heatmaps showing the mean and the standard deviations created with GraphPad Prism

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Cell-penetrating peptides (CPPs), referred to as protein transduction domains (PTDs) or Trojan peptides, form a structurally diverse group of molecules with varied physicochemical properties. They usually have at least two features in common. They can penetrate the cell membrane barrier even after binding with macromolecules. They are mostly short peptides with a length of up to 30 amino acids [1]. In vitro and in vivo studies have indicated their low cytotoxicity and immunogenicity together with high efficiency

Methods

Results

Discussion

Conclusion