Cell‐penetrating peptide–labelled smart polymers for enhanced gene delivery

Abstract

Highly efficient gene delivery vehicles are pursued to progress gene therapy. In this study, we developed the cell‐penetrating peptide‐labelled and degradable gene carriers for efficient external gene transfection. The cationic carriers were prepared by coupling low‐molecular‐weight polyethylenimine (PEI800) with 4ʹ4‐dithiodibutyric acid (DA), and HIV‐1 Trans‐Activator of Transcription (TAT) was conjugated to the carriers as a penetrating peptide. The resulted PEI‐DA‐TAT was able to condense plasmid DNA (pDNA) into a complex with a hydrodynamic size of around 150 nm under a neutral condition. PEI‐DA‐TAT showed negligible cytotoxicity to both Hela and HEK 293 cells with the cell viability of more than 80% beyond the carrier concentration of 50 μg/mL. This new carrier displayed better performance in regard to DNA transfection efficiency in comparison with the carriers of non‐TAT labelled PEI‐DA, commercial PEI25K and low‐molecular‐weight PEI (PEI800). The transfection efficiency of PEI‐DA‐TAT was increased by 8% compared with PEI‐DA and PEI25K. The experimental findings suggested that the developed PEI‐DA‐TAT is a promising carrier for efficient DNA delivery with low cytotoxicity for gene therapy.

Gene delivery by peptide-assisted transport

“…Meanwhile, CPP-triggered endocytosis is driven by CPP binding to membrane components, leading to energy- and temperature-dependent membrane invagination and uptake. For example, a study by Zhang et al utilized HIV-1 TAT conjugated to PEI800 with 4’4-dithiodibutyric acid (DA) to develop CPP-labelled degradable gene carriers for efficient gene transfection (PEI-DA-TAT) [27]. These studies showed that the cationic HIV-1 TAT peptide interacted with the negatively charged cell membrane and thereby stimulated endocytosis of the peptide along with the gene carrier system.…”

Nanocarrier-based gene delivery for immune cell engineering

“…[161][162][163][164] In recent works, peptides like TAT, GALA and KALA were used to induce endosomal escape by enhancing the proton buffering sponge effect. 52,[165][166][167] Interestingly, histidine-rich peptides attached to poly(amidoamine) (PAMAM) dendrimers showed a significant effect on enhanced ability of endosomal escape through the balloon puncturing effect (proton sponge effect + endosomal destabilization). 168 4.1.4 Efficiency of nuclear internalization.…”

Self-Assembled Biodegradable Core-Shell Nanocomposites of Amphiphilic Retinoic Acid-LMW bPEI Conjugates Exhibit Enhanced Transgene Expression in Hepatocellular Carcinoma Cells With Inherent Anticancer Properties

“…7,8 Some successful attempts have been made to reduce its toxicity with simultaneous increase in the transfection efficiency, still, systemic clearance remains as an unaddressed issue. To mimic the molecular weight and structural features of bPEI (25 kDa), low molecular weight bPEI has been engineered in several ways, viz., by crosslinking (covalent or stimuli responsive), 9 attaching targeting ligands, 10 tethering dendrimers, 11 introducing hydrophobic ligands, 12 conjugating cell penetrating peptides, 13 grafting polysaccharides, 14 polyethylene glycols, 15 etc. Structurally, bPEI, being a hydrophilic polymer, lacks hydrophobic domains which have now been demonstrated to play an important role during the course of interactions with the cell membrane.…”