Abstract
BackgroundCell penetrating peptides (CPPs) as one class of non-viral vectors, have been widely explored as a delivery tool due to their cell-penetrating capability with low cytotoxicity. However, CPPs have reported to have low gene transfection efficiency mainly due to the fact that DNA is larger than other biomolecules. On the other hand, the conventional linear CPPs are unstable for constructing the DNA complexes with it. Thus, here we designed a branched CPP using disulfide bridges based on the linear TAT peptide, to enhance the gene delivery efficiency in a better way.ResultsThe branched TAT (BTAT) was synthesized by the DMSO oxidation method and showed high-molecular-weight about 294 kDa. The resulting BTAT was complexed with plasmid green fluorescence protein (pGFP) gene at various N/P ratios. The gene transfection efficiency was assessed on HeLa cells after treating with BTAT/pGFP complexes, showed high gene transfection efficiency as conformed by flowcytometry followed by confocal laser scanning microscopy (CLSM) visualization.ConclusionThe novel BTAT/pGFP complex exhibited significantly higher stability and redox cleavability by reducing agent. In addition, BTAT showed higher transfection efficiency approximately 40-fold than those of the TAT and mTAT complexes. Our primary experiments demonstrated the potential of BTAT as a suitable candidate for gene delivery and it could be applied for various types of gene delivery platforms.
Highlights
Cell penetrating peptides (CPPs) as one class of non-viral vectors, have been widely explored as a delivery tool due to their cell-penetrating capability with low cytotoxicity
After dimethyl sulfoxide (DMSO) oxidation, the Modified TAT (mTAT) solution was changed to clear gel form, indicative of branched TAT (BTAT) formation
After DMSO oxidation, the absolutemolecular-weight was measured by static light scattering (SLS) using the various concentrations of BTAT
Summary
Cell penetrating peptides (CPPs) as one class of non-viral vectors, have been widely explored as a delivery tool due to their cell-penetrating capability with low cytotoxicity. Here we designed a branched CPP using disulfide bridges based on the linear TAT peptide, to enhance the gene delivery efficiency in a better way. Gene therapy is a powerful tool with the potential to inhibit the deleterious effects of malicious genes by inserting corrected/normal genes into the genome to treat the disease It could treat the disease by delivering specific nucleic acid into the target cells instead of drugs [1]. For over 20 years, CPP based gene delivery has been investigated [17,18,19,20,21,22,23] to enhance the transfection efficiency and introduce the targeting capability to the CPP/DNA complexes. The first CPPs were truncated from the transduction domain of the HIV-virus, TAT (48–60), since it has been widely studied [24]
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