Department of General Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, 141/143 Pomorska St., 90-236 Lodz, Poland.
Institute of Macromolecular Compounds, Russian Academy of Sciences, Bolshoi Prospect 31, V.O., 199004 St. Petersburg, Russia.
Int J Mol Sci. 2020 Apr 29;21(9):3138. doi: 10.3390/ijms21093138.
The disruption of the cellular pathways of protein biosynthesis through the mechanism of RNA interference has been recognized as a tool of great diagnostic and therapeutic significance. However, in order to fully exploit the potential of this phenomenon, efficient and safe carriers capable of overcoming extra- and intracellular barriers and delivering siRNA to the target cells are needed. Recently, attention has focused on the possibility of the application of multifunctional nanoparticles, dendrimers, as potential delivery devices for siRNA. The aim of the present work was to evaluate the formation of dendriplexes using novel poly(lysine) dendrimers (containing lysine and arginine or histidine residues in their structure), and to verify the hypothesis that the use of these polymers may allow an efficient method of siRNA transfer into the cells in vitro to be obtained. The fluorescence polarization studies, as well as zeta potential and hydrodynamic diameter measurements were used to characterize the dendrimer:siRNA complexes. The cytotoxicity of dendrimers and dendriplexes was evaluated with the resazurin-based assay. Using the flow cytometry technique, the efficiency of siRNA transport to the myeloid cells was determined. This approach allowed us to determine the properties and optimal molar ratios of dendrimer:siRNA complexes, as well as to demonstrate that poly(lysine) dendrimers may serve as efficient carriers of genetic material, being much more effective than the commercially available transfection agent Lipofectamine 2000. This outcome provides the basis for further research on the application of poly(lysine) dendrimers as carriers for nucleic acids in the field of gene therapy.
通过 RNA 干扰机制破坏蛋白质生物合成的细胞途径已被认为是一种具有重要诊断和治疗意义的工具。然而，为了充分利用这一现象的潜力，需要能够克服细胞内外障碍并将 siRNA 递送到靶细胞的高效和安全载体。最近，人们关注的焦点是多功能纳米粒子（如树枝状聚合物）作为 siRNA 潜在递送载体的应用可能性。本工作的目的是评估使用新型聚（赖氨酸）树枝状聚合物（其结构中含有赖氨酸和精氨酸或组氨酸残基）形成树枝状聚合物-核酸复合物，并验证使用这些聚合物可能获得将 siRNA 有效递送到细胞中的方法的假设。荧光偏振研究以及 zeta 电位和水动力直径测量用于表征树枝状聚合物：siRNA 复合物。使用基于 Resazurin 的测定法评估树枝状聚合物和树枝状聚合物-核酸复合物的细胞毒性。通过流式细胞术技术测定 siRNA 向髓样细胞的转运效率。这种方法使我们能够确定树枝状聚合物：siRNA 复合物的性质和最佳摩尔比，并证明聚（赖氨酸）树枝状聚合物可以作为遗传物质的有效载体，其效果远优于市售转染试剂 Lipofectamine 2000。这一结果为进一步研究聚（赖氨酸）树枝状聚合物作为基因治疗领域中核酸载体的应用提供了基础。
