Targeting of Cellular Organelles by Fluorescent Plasmid DNA Nanoparticles

Abstract

The development of a suitable delivery system and the targeting of intracellular organelles are both essential for the success of drug and gene therapies. The conception of fluorescent ligands, displaying targeting specificity together with low toxicity, is an emerging and reliable tool to develop innovative delivery systems. Biocompatible BSA or pDNA/ligand nanoparticles were synthesized by a coprecipitation method and were shown to display adequate sizes and morphology for delivery purposes, and positive surface charges. Additionally, these fluorescent vectors can target specific intracellular organelles. In vitro transfection mediated by BSA or pDNA based carriers can result in the accumulation of BSA in the cytosol, lysosomes, and mitochondria or the expression of the plasmid-encoded protein, respectively. Moreover, the therapeutic effect of pDNA/ligand vectors in cancer gene therapy instigates further research aiming clinical translation.

The Performance of Minicircle DNA Versus Parental Plasmid in p53 Gene Delivery Into HPV-18-Infected Cervical Cancer Cells

“…In fact, the results mentioned above have suggested that the mcDNA-p53 vector is effective in decreasing cancer cell viability, which can be associated with this repression of cell proliferation. This growth inhibitory effect was also found in a similar assay performed by Costa et al, after transfection with a pDNA vector encoding the p53 gene [28]. Thus, it can be concluded that overexpression of p53 protein by mcDNA can lead to the reinstatement of p53 functions, which was once affected by E6 degradation mechanisms.…”

Development of Peptide-Based Nanoparticles for Mitochondrial Plasmid DNA Delivery

“…In the last decades, considerable advances have been made in the conception of nucleic acid-based delivery systems to overcome the major drawbacks of payload delivery to eukaryotic cells, namely, cellular uptake/internalization, endosomal escape, targeting a specific subcellular compartment, and ultimately, the induction of therapeutic action [19,[21][22][23][24]. In line with this aim, micelles, polymers, lipid-and peptide-based nanoparticles are among the most studied systems for gene release [25][26][27][28].…”

Delivery Systems for Mitochondrial Gene Therapy: A Review

“…To confirm the affinity of the systems for mitochondria, after cellular transfection, mitochondria were separated from the cytosolic fraction, verifying greater accumulation of nanoparticles in isolated mitochondria compared to the cytosol fraction, through quantification of rhodamine fluorescence [ 145 ]. Costa et al also created calcium carbonate delivery systems capable of encapsulating p53 and ND1-GFP plasmids [ 146 ]. The nanoparticles formulated with the compound [16]phenN 2 showed an affinity for mitochondria.…”

“…The fluorescence of [16]phenN 2 in isolated mitochondria was quite high when compared to the fluorescence of this compound in the cytosol or in lysosomes, where fluorescence was residual. Mitochondrial targeting of systems with [16]phenN 2 was further reinforced when plasmids were replaced by BSA, which resulted in an accumulation of BSA within the mitochondria of cells transfected with these nanoparticles, while this accumulation did not occur in the cytosol and the lysosome [ 146 ].…”