Effect of Degree of Substitution and Molecular Weight on Transfection Efficacy of Starch-Based siRNA Delivery System
Abstract
:1. Introduction
2. Materials and Methods
2.1. Starch Quaternization
2.2. Quaternized Starch Chemical Analysis
2.3. Q-Starch(DS)/siRNA Complex Preparation
2.4. Q-Starch(DS)/siRNA Complex Characterization
2.4.1. Agarose Gel Electrophoresis
2.4.2. Dynamic Light Scattering (DLS)
2.4.3. Ionic Strength Agarose Gel Electrophoresis
2.4.4. Zeta Potential
2.4.5. Cryo-Transmission Electron Microscopy (cryoTEM)
2.5. Cell Culture Handling
2.6. In Vitro Cellular Uptake
2.7. siRNA: EGFR Gene Silencing
2.8. Iodine Starch Test
2.9. Viscosity Measurements
2.10. Statistical Analysis
3. Results and Discussion
3.1. Q-Starch Synthesis and Chemical Characterization
3.2. Q-Starch(DS)/siRNA Complex Formation and Characterization
3.3. Q-Starch(DS)/siRNA Electrostatic Strengths and Size Distributions
3.4. Q-Starch(DS)/siRNA Cellular Uptake and Gene Silencing Capabilities
3.5. Enzymatic Cleavage of Q-Starch(DS) by α-Amylase
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Q-Starch Average Nitrogen Content (wt. %) | DS * | (DS)−1 |
---|---|---|
0.38 ± 0.00 | 0.05 | 21.71 |
0.63 ± 0.00 | 0.08 | 12.87 |
0.93 ± 0.07 | 0.12 | 8.39 |
2.03 ± 0.08 | 0.30 | 3.32 |
2.68 ± 0.09 | 0.44 | 2.29 |
3.3 ± 0.1 | 0.59 | 1.70 |
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Regev, A.; Benafsha, C.; Goldbart, R.; Traitel, T.; Elkabets, M.; Kost, J. Effect of Degree of Substitution and Molecular Weight on Transfection Efficacy of Starch-Based siRNA Delivery System. Polysaccharides 2024, 5, 580-597. https://doi.org/10.3390/polysaccharides5040037
Regev A, Benafsha C, Goldbart R, Traitel T, Elkabets M, Kost J. Effect of Degree of Substitution and Molecular Weight on Transfection Efficacy of Starch-Based siRNA Delivery System. Polysaccharides. 2024; 5(4):580-597. https://doi.org/10.3390/polysaccharides5040037
Chicago/Turabian StyleRegev, Amir, Chen Benafsha, Riki Goldbart, Tamar Traitel, Moshe Elkabets, and Joseph Kost. 2024. "Effect of Degree of Substitution and Molecular Weight on Transfection Efficacy of Starch-Based siRNA Delivery System" Polysaccharides 5, no. 4: 580-597. https://doi.org/10.3390/polysaccharides5040037