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. 2017 Aug 22;7(1):9159.
doi: 10.1038/s41598-017-09803-z.

Magnetic Nanoparticle Assisted Self-assembly of Cell Penetrating Peptides-Oligonucleotides Complexes for Gene Delivery

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Free PMC article

Magnetic Nanoparticle Assisted Self-assembly of Cell Penetrating Peptides-Oligonucleotides Complexes for Gene Delivery

Moataz Dowaidar et al. Sci Rep. .
Free PMC article

Abstract

Magnetic nanoparticles (MNPs, Fe3O4) incorporated into the complexes of cell penetrating peptides (CPPs)-oligonucleotides (ONs) promoted the cell transfection for plasmid transfection, splice correction, and gene silencing efficiencies. Six types of cell penetrating peptides (CPPs; PeptFect220 (denoted PF220), PF221, PF222, PF223, PF224 and PF14) and three types of gene therapeutic agents (plasmid (pGL3), splicing correcting oligonucleotides (SCO), and small interfering RNA (siRNA) were investigated. Magnetic nanoparticles incorporated into the complexes of CPPs-pGL3, CPPs-SCO, and CPPs-siRNA showed high cell biocompatibility and efficiently transfected the investigated cells with pGL3, SCO, and siRNA, respectively. Gene transfer vectors formed among PF14, SCO, and MNPs (PF14-SCO-MNPs) showed a superior transfection efficiency (up to 4-fold) compared to the noncovalent PF14-SCO complex, which was previously reported with a higher efficiency compared to commercial vector called Lipofectamine™2000. The high transfection efficiency of the new complexes (CPPs-SCO-MNPs) may be attributed to the morphology, low cytotoxicity, and the synergistic effect of MNPs and CPPs. PF14-pDNA-MNPs is an efficient complex for in vivo gene delivery upon systemic administration. The conjugation of CPPs-ONs with inorganic magnetic nanoparticles (Fe3O4) may open new venues for selective and efficient gene therapy.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Schematic representation of CPP-ONs-MNPs noncovalent complexes. The size of the objects is not considered.
Figure 2
Figure 2
Characterization of MNPs using (a) XRD, (b) TEM, and (c) zeta potential.
Figure 3
Figure 3
SEM images of (a) PF221-pGL3-MNPs, (b) PF220-SCO-MNPs and (c) PF14-siRNA-MNPs. The insets in (a) and (b) are enlarged areas showing the subunits of the nanocomplexes.
Figure 4
Figure 4
TEM images of (a) PF220-pGL3-MNPs, (b) PF221-pGL3-MNPs, (c) PF14-pGL3-MNPs), (d) PF220-SCO-MNPs, (e) PF221-SCO-MNPs, (f) PF14-SCO-MNPs, and PF14-siRNA-MNPs for siRNA of 25 nM (g) and 50 nM (h).
Figure 5
Figure 5
Transfection of the pGL3 plasmid in Hela cells for CPPs-pGL3 (CR5) and CPPs-pGL3-MNPs (CR5 + Fe3O4); while CPPs are PF220, PF221, PF222, PF223, and PF224 using a charge ratio of 5 (CR5), RLU refers to the relative light unit (RLU).
Figure 6
Figure 6
Splice-correction activity of (a) CPPs-SCO and CPPs-SCO-MNPs (CPPs of PF220, PF221, PF222, PF223 and PF224) and (b) PF14-SCO complexes and PF14-SCO-MNPs with and without purification.
Figure 7
Figure 7
(a) PF14 and PF14-MNPs mediate efficient siRNA in U87 Luc cell-lines, (b) cell uptakes based on the mechanism of scavenger receptor class, (c,d) confocal microscopy images of Hela-705 cells incubated for 24 h with PF14-Alexa 568–705ASO (MR10, 100 nM ASO) complexes (in red) without (c) and with (d) MNPs. Cell-membranes are stained with Fast-dio stain (in green).
Figure 8
Figure 8
In vivo gene delivery efficacies of PF14 and PF14-pDNA-MNPs at N/P4 using pLuc. The data are represented as a fold increase of RLU/mg over the treatment with pDNA. N represents 2–5 repeated experiments.

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