Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. Preparation of siRNA/MCD Complex
Group | Name | Basic structure | R (L-chain) |
---|---|---|---|
Type-C | C-1 | -C14H25 | |
C-2 | -C16H33 | ||
Type-D | D-1 | -C12H25 | |
D-2 | -C16H33 | ||
Type-E | E-1 | -C12H25 | |
E-2 | -C14H29 | ||
E-3 | -C16H33 |
2.2. Comparison of Gene Silencing Efficiencies and Cell Viabilities Between siRNA/MCD Nanoparticles and LFN 2000
2.3. Investigation of siRNA Nanoparticle Formed with Various Type of MCD on Gene Silencing Efficiencies
MCD (N/P ratio) | siRNA/MCD nanoparticle | R8-D-MEND | ||||
---|---|---|---|---|---|---|
Size (nm) | Zeta potential (mV) | PDI | Size (nm) | Zeta potential (mV) | PDI | |
C-1 (1.0) | 130 ± 65 | −15.5 ± 19.3 | 0.48 ± 0.08 | 111 ± 9 | 32.4 ± 3.6 | 0.22 ± 0.04 |
C-2 (5.0) | 131 ± 60 | 13.1 ± 5.6 | 0.35 ± 0.11 | 110 ± 3 | 43.3 ± 3.1 | 0.24 ± 0.06 |
D-1 (5.0) | 51 ± 18 | −13.1 ± 7.7 | 0.32 ± 0.15 | 92 ± 13 | 30.3 ± 13.4 | 0.20 ± 0.03 |
D-2 (5.0) | 85 ± 27 | 10.2 ± 3.9 | 0.25 ± 0.02 | 123 ± 10 | 45.0 ± 3.6 | 0.18 ± 0.03 |
E-1 (2.5) | 85 ± 26 | −34.6 ± 11.4 | 0.28 ± 0.08 | 105 ± 9 | 30.9 ± 6.4 | 0.20 ± 0.05 |
E-2 (7.5) | 60 ± 8 | 4.9 ± 3.4 | 0.24 ± 0.03 | 109 ± 11 | 43.7 ± 0.8 | 0.20 ± 0.03 |
E-3 (5.0) | 67 ± 7 | 32.6 ± 2.5 | 0.33 ± 0.06 | 113 ± 7 | 51.0 ± 2.5 | 0.26 ± 0.02 |
3. Experimental
3.1. Materials
3.2. Preparation of siRNA/MCD Nanoparticle
3.3. Measurement of Size and Zeta-Potential of Carriers
3.4. Construction of R8-D-MEND Containing siRNA/MCD Nanoparticle
3.5. Evaluation of Gene Silencing Efficiency and Cell Viability
3.6. Statistical Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Supplementary Materials
Nanoparticles prepared using C-1 | Nanoparticles prepared using C-2 | ||||||
---|---|---|---|---|---|---|---|
N/P ratio | Size (nm) | Zeta potential (mV) | PDI | N/P ratio | Size (nm) | Zeta potential (mV) | PDI |
0.25 | 94 | −37.9 | 0.72 | 0.25 | 162 | −42.9 | 0.63 |
0.5 | 124 | −29.8 | 0.55 | 0.5 | 2520 | −19.9 | 0.37 |
1 | 66 | −28.8 | 0.33 | 2.5 | 616 | 1.6 | 0.6 |
1.5 | 90 | −37.5 | 0.36 | 3.75 | 584 | 10.7 | 0.57 |
2 | 90 | −28.6 | 0.26 | 5 | 151 | 18.7 | 0.43 |
2.5 | 1480 | −28.1 | 0.58 | 6.25 | 108 | 19.9 | 0.27 |
5 | 1650 | −9.3 | 0.39 | 7.5 | 58 | 27.7 | 0.37 |
10 | 1360 | 0.8 | 0.56 | 8.75 | 84 | 29.3 | 0.42 |
10 | 57 | 31.7 | 0.36 | ||||
Nanoparticles prepared using D-1 | Nanoparticles prepared using D-2 | ||||||
N/P ratio | Size (nm) | Zeta potential (mV) | PDI | N/P ratio | Size (nm) | Zeta potential (mV) | PDI |
0.25 | 244 | −5.6 | 0.67 | 0.25 | 156 | −47.8 | 0.44 |
0.5 | 302 | −19.9 | 0.35 | 0.5 | 2560 | −18.6 | 0.7 |
2.5 | 162 | −18.5 | 0.34 | 2.5 | 888 | 4.5 | 0.62 |
3.75 | 113 | −34.4 | 0.34 | 3.75 | 69 | 18.3 | 0.24 |
5 | 123 | −37.0 | 0.22 | 5 | 95 | 18.7 | 0.32 |
6.25 | 69 | −35.0 | 0.3 | 6.25 | 82 | 21 | 0.34 |
7.5 | 55 | −31.9 | 0.21 | 7.5 | 65 | 30.4 | 0.36 |
8.75 | 99 | −33.5 | 0.24 | 8.75 | 50 | 30 | 0.34 |
10 | 102 | −35.0 | 0.25 | 10 | 53 | 31.3 | 0.36 |
Nanoparticles prepared using E-1 | Nanoparticles prepared using E-2 | ||||||
N/P ratio | Size (nm) | Zeta potential (mV) | PDI | N/P ratio | Size (nm) | Zeta potential (mV) | PDI |
0.25 | 713 | −19.9 | 0.85 | 0.25 | 133 | −50.2 | 0.41 |
0.5 | 198 | −20.0 | 0.49 | 0.5 | 129 | −29.6 | 0.24 |
1.25 | 324 | −20.7 | 0.66 | 2.5 | 3140 | −9.3 | 0.455 |
2.5 | 74 | −34.2 | 0.28 | 3.75 | 918 | 0.3 | 0.45 |
3.75 | 159 | −29.1 | 0.18 | 5 | 221 | 8.2 | 0.386 |
5 | 165 | −28.5 | 0.14 | 6.25 | 132 | 12.9 | 0.22 |
7.5 | 2030 | −20.5 | 0.7 | 7.5 | 53 | 16.5 | 0.26 |
10 | 2160 | −18.8 | 0.6 | 8.75 | 54 | 16.9 | 0.28 |
10 | 63 | 20.5 | 0.313 | ||||
Nanoparticles prepared using E-3 | |||||||
N/P ratio | Size (nm) | Zeta potential (mV) | PDI | ||||
2.5 | 54 | 25.2 | 0.27 | ||||
3.75 | 63 | 40.1 | 0.25 | ||||
5 | 53 | 39.7 | 0.36 | ||||
7.5 | 45 | 44.4 | 0.32 | ||||
10 | 57 | 42.8 | 0.42 |
Size (nm) | Zata potential (mV) | |
---|---|---|
SUV | 74 ± 10 | −66.8 ± 8.9 |
R8-SUV | 86 ± 14 | 29.9 ± 11.3 |
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Yamada, Y.; Suzuki, R.; Harashima, H. Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells. Cancers 2013, 5, 1413-1425. https://doi.org/10.3390/cancers5041413
Yamada Y, Suzuki R, Harashima H. Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells. Cancers. 2013; 5(4):1413-1425. https://doi.org/10.3390/cancers5041413
Chicago/Turabian StyleYamada, Yuma, Ryosuke Suzuki, and Hideyoshi Harashima. 2013. "Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells" Cancers 5, no. 4: 1413-1425. https://doi.org/10.3390/cancers5041413