Hydrophobic and electrostatic interactions between cell penetrating peptides and plasmid DNA are important for stable non-covalent complexation and intracellular delivery

Abstract

Cell penetrating peptides are useful tools for intracellular delivery of nucleic acids. Delivery of plasmid DNA, a large nucleic acid, poses a challenge for peptide mediated transport. The paper investigates and compares efficacy of five novel peptide designs for complexation of plasmid DNA and subsequent delivery into cells. The peptides were designed to contain reported DNA condensing agents and basic cell penetrating sequences, octa-arginine (R ) and CHK HC coupled to cell penetration accelerating peptides such as Bax inhibitory mutant peptide (KLPVM) and a peptide derived from the Kaposi fibroblast growth factor (kFGF) membrane translocating sequence. A tryptophan rich peptide, an analogue of Pep-3, flanked with CH on either ends was also a part of the study. The peptides were analysed for plasmid DNA complexation, protection of peptide-plasmid DNA complexes against DNase I, serum components and competitive ligands by simple agarose gel electrophoresis techniques. Hemolysis of rat red blood corpuscles (RBCs) in the presence of the peptides was used as a measure of peptide cytotoxicity. Plasmid DNA delivery through the designed peptides was evaluated in two cell lines, human cervical cancer cell line (HeLa) and (NIH/3 T3) mouse embryonic fibroblasts via expression of the secreted alkaline phosphatase (SEAP) reporter gene. The importance of hydrophobic sequences in addition to cationic sequences in peptides for non-covalent plasmid DNA complexation and delivery has been illustrated. An alternative to the employment of fatty acid moieties for enhanced gene transfer has been proposed. Comparison of peptides for plasmid DNA complexation and delivery of peptide-plasmid DNA complexes to cells estimated by expression of a reporter gene, SEAP. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

The Effect of Conjugation with Octaarginine, a Cell-Penetrating Peptide on Antifungal Activity of Imidazoacridinone Derivative

“…In effect, the DNA-peptide complex was immobile during electrophoresis and remained in wells. Similar results were reported in studies on cell-penetrating peptides and DNA complexation [44].…”

Dual-Modified Liposome for Targeted and Enhanced Gene Delivery into Mice Brain

“…Mel derivatives have shown to enhance intracellular delivery of therapeutic macromolecules (Kyung et al, 2018). Hydrophobic sequences such as Kaposi fibroblast growth factor (kFGF) have been reported relevant for intracellular delivery of nucleic acids, providing stable noncovalent DNA complexation and protection against nucleases together with the ability to translocate across the lipid bilayer of plasma membrane (Lin et al, 1995;Upadhya and Sangave, 2016;Bolhassani et al, 2017). The addition of hydrophobic sequence FFLIPKG, known as penetration accelerating sequence (Pas), to the arginine-rich peptide R8 formed the hybrid PasR8, revealed not only to improve the carrier abilities but also to facilitate the escape from endocytic lysosomes (Takayama et al, 2009(Takayama et al, , 2012.…”

Prospect of cell penetrating peptides in stem cell tracking

“…Covalent bonding requires unique conjugation design for different cargoes, so non-covalent bonding is more widely used. Non-covalent bonding include biotin–streptavidin interaction [ 103 ], electrostatic Interactions [ 57 ], and metal affinity interactions [ 103 ]. Although the non-covalent binding method is more versatile, it is still difficult for biological laboratories that lacking a chemical background to synthesize CPPs-modified imaging nanoparticles.…”

“…There are two ways to conjugate CPPs to the cargo: covalent and non-covalent binding [ 22 ] Although both methods are feasible, in practical applications, the second method is more popular [ 57 ]. In non-covalent approach, CPPs bind to cargo through electrostatic interaction [ 29 ].…”