Table 1.
Structure Characterization | Biological activity | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
N° | DNA Nanostructure | Characterization | Size (nm) | Cargo (Loading) | Stability | Targeting | Loading efficiency | Release | Internalization | In vitro tests | In vivo tests | Biodistribution | Ref | ||
Cells | Activity | Cells | Activity | ||||||||||||
1 | Tetrahedral | AFM DLS PAGE (6%) | AFM: height 2-3 nm DLS: 9.08 nm ± 3nm | Doxo | > 4 < 23 hrs Solution: 10% human serum Temp: 37°C | N/A | 26 molecules/structure Time: 1 hr Temp: N/A | 100% in 10hrs. <10% in 3hrs. Solution: PBS pH 7.4 Temp: 37°C 60% in 10 hrs Solution: PBS pH 5 Temp: 37°C | Macropinocytosis and caveolae-mediated endocytosis pathways | Breast cancer MCF7 (drug sensitive) and MCF7-ADR (Doxo resistant). | Doxo-tetrahedral is more effective on cell viability of MCF7-ADR than free doxo. Same efficacy on MCF7 cells. | N/A | N/A | N/A | [63] |
2 | Icosahedral: five- (120 bp) and six- (144 bp) (aptamer) point-star structure | AGE (2%) DLS TEM | DLS: - Five stars (28.2 nm ± 3nm) - Six stars (28.6 nm ± 5nm). TEM: 25 nm | Doxo | >30min Solution: cell culture medium Temp: 37°C | Mucin 1 (MUC1). Tumor surface marker | 1200 molecules/structure Time: 1h incubation Temp: room temperature | N/A | Dynamin-dependent and clathrin-mediated endocytosis. Degradation in lysosomes | Breast cancer MCF7 (MUC1+), CHO-K1 (MUC1-) | Doxo-Aptamer six-point-star structure is more effective on cell viability of MUC1+ cell than free doxo. Same efficacy on MUC- cells. | N/A | N/A | N/A | [64] |
3 | Tetrahedral (30bp/edge, Total 6 edges) | AFM DLS PAGE (5%) | AFM: height 7.5 nm. DLS: 28.6 nm ± 2.38 nm | siRNA against GFP gene. DNA structure conjugated with folic acid. | N/A | Folic acid receptor (FAR) | 1-6 siRNA per tetrahedral (1 siRNa/edge) | N/A | N/A | HeLa cells (LUC+). KB cells (HeLa cell contaminant overexpressing folate receptor) | HeLa: LUC expression < 50%. KB: GFP expression <40% | KB xenograft tumours | siRNA no effect on LUC. siRNA-DNA- tetrahedral: IC 50 LUC expression: 1.8 mg/kg. Inj: intratumor or tail-vein Stability: siRNA 6 min; siRNA-DNA tetrahedral 24 min. | Tumor, Kidney Time: 12 hours post injection | [65] |
4 | Tetrahedral | PAGE (3.5%); | N/A | Straptavidin (STV) and CpGs | 5hrs Solution: FBS 50% Temp: room temperature | N/A | N/A | N/A | Endocytosis. Antigen localized in lysosomes after 2 hours | RAW264.7 (macrophage, Abelson murine leukemia virus transformed). | Increased internalization of the complex tetrahedron-STV-CpG by APC cells | BALB/c immunocompetent mice | Mice immunized with the complex tetrahedron-STV-CpG developed a stronger and a longer immunitary response. | N/A | [66] |
5 | Aptamer-tethered DNA nanotrain | AFM AGE (3%) TEM | AFM: length 100 nm. | Doxo, epirubicin, daunorubicin | >45 hrs Solution: PBS + 5mM Mg2+ Temp: N/A | Protein Tyrosin Kinase 7 (PTK7) | N/A | N/A | Endocytosis | CEM cells (human T-cell acute lymphocytic leukemia PTK7+) and Ramos (human B lymphocyte Burkitt's lymphoma PTK7-) | Drug-Aptamer-DNA-drug is more cytotoxic on PTK7+ cells than free drug. Same efficacy on PTK7- cells. | CEM (PTK7+) xenograft mouse model | Increased antitumor efficacy and reduced side effects of doxo delivered via Aptamer-DNA nanotrain. Inj: I.V. | [67] | |
6 | Open Caged DNA (pyramidal), 408 bp | PAGE (7%) | N/A | Doxo | 35 hrs (half-life) Solution: cell culture medium+10% FBS Temp: 37°C | N/A | 172 molecules doxo/structure (at 15% of loading efficiency) | 50% of Doxo release from py-Doxo in PBS in 5 hrs and 3 hrs in FBS. Free Doxo in 20 minutes | py-Doxo is able to penetrate inside MDA-MB-231, release Doxo in the nucleus | MDA-MB-231, HepG2 | Decrease cell viability compared to free doxo | N/A | N/A | N/A | [68] |
7 | Tube with different global twist (Straight (S) 10.5 bp/turn and Twist (T) tube 12 bp/turn) | AGE (2%) TEM | TEM: length: 138 nm, diameter: 13 nm | Doxo | 48hr (T-tube) Solution: 10% FBS Temp: 37°C | N/A | N/A | 80% (T-tube) and 90% (S-tube) in 10 hrs Solution: PBS pH 7.4 Temp: 37°C | N/A | Breast cancer MDA-MB-231; MDA-MB-468; MCF-7 | In all the cell lines tested the T-DNA IC50 is ≈2 times lower than free Doxo. (calculated by our group) | N/A | N/A | N/A | [70] |
8 | Tube | AGE (2%) TEM | TEM: length: 80 nm, diameter: ≈20 nm | 62 CpG sequence specific for mouse Toll like receptor 9 | 6 hrs Solution: cell culture medium Temp: 37°C | Toll like receptor 9 (TLR9). | 62 binding sites per tube | N/A | N/A | Splenocytes from female C57BL/6 mice | N/A Immunoresponse through the TLR9. Nontoxic | N/A | N/A | N/A | [71] |
9 | Hexagonal barrel with aptamer-based lock (antigen keys). | AFM AGE (2%) DLS TEM | DLS: 90 nm TEM: 35nm x 35 nm x 45 nm | Fab antibody fragments | N/A | CD33, CDw328 | N/A | N/A | N/A | NKL | Increase apoptosis | N/A | N/A | N/A | [72] |
10 | Triangle and tube | AFM AGE (1%) | AFM: -Triangle: edge ≈ 150nm -Tube: length ≈ 183 nm | Doxo | N/A | N/A | >200000 molecules (calculated by our group) Time: 24 hrs Temp: room temperature. | ≈15% pH 7.4, 35% pH 5.5 in 48 hrs ≈25% in MCF7 cell lysate >40% with 50U DNAaseI Solution: PBS Temp: 37°C | Endocytosis and localization of origami in lysosomes after 6 hours of treatment. | Breast cancer MCF7 and MCF7 resistant | MCF-7: 2.5µM and MCF-7 resistant >100μM. DNA origami loaded with doxo enhanced the cells death compared to free doxo on MCF7 doxo resistant cells. No differences between free doxo and origami doxo in regular MCF7. | N/A | N/A | N/A | [73] |
11 | Tube, triangle and square | AFM AGE (1%) DLS | AFM: -Tube: height: 7 nm, diameter: 380 nm, -Triangle: edge 120 nm, -Square: length*width 90nmx60nm DLS: -Triangle: 59 nm -Square: 80.9 nm -Tube: 98.6 nm. | Doxo | 24 hrs Solution: serum Temp: 37°C | N/A | >200000 molecules (calculated by our group) Time: 24 hrs Temp: room temperature. | ≈20% of Doxo is released in 48 hours at pH 7.4, ≈35% is released at pH 5.5 Solution: PBS Temp: 37°C | N/A | Breast cancer MDA-MB-231 | No significant difference compared to the free doxo. | MDA-MB-231 cells Orthotopic breast cancer model | Increased EPR effect of DNA origami. Significant tumor reduction in mice treated with doxo-origami compared to free doxo Inj: I.V | Tumor: Triangle > tube >square Tumor > liver > kidney > spleen Time: 24 hrs | [74] |
12 | Tube (Tile) | AGE (2%) TEM | TEM: Length: 27 nm and diameter: 8 nm | siRNA (GFP) | 8hrs Solution: cell culture medium Temp: 37 °C (Degradation depends on Mg2+ concentration, oligonucleotide sequences, salt concentration, structural extension) | Folic acid receptor (FAR) | N/A | N/A | Endosomal trapping (no release) | N/A | No effects on GFP expression | N/A | NA | NA | [86] |
AGE: Agarose gel electrophoresis. PAGE: Polyacrylamide gel electrophoresis. TEM: Transmission electron microscopy. DLS: Dynamic light scattering. AFM: Atomic force microscopy. N/A: Not Applicable. Temp: Temperature. Inj: Injection. I.V.: Intravenous. Luc: Luciferase