Ho Uyen, El-Bakkari Mounir, Alshamsan Aws, Cho Jae-Young, Yamazaki Takeshi, Hemraz Usha D, Fenniri Hicham
Department of Chemistry, University of Alberta, 11227 Saskatchewan Drive Edmonton, Alberta, T6G 2G2, Canada.
Nanotechnology Research Centre, National Research Council of Canada, 11421 Saskatchewan Drive, Edmonton, Alberta, T6G 2M9, Canada.
Biomater Sci. 2023 Oct 24;11(21):7169-7178. doi: 10.1039/d3bm01115a.
The quest for new therapeutic treatments for hereditary diseases has led to many advances in RNA interference (RNAi) and gene silencing. While this technique has the potential to address many problems, the key to its continued use is the development of effective delivery strategies that would reduce cellular toxicity and increase silencing efficiency. Rosette nanotubes (RNTs) are biomimetic supramolecular nanostructures formed through the self-assembly of hybrid guanine-cytosine (G∧C) DNA bases. Here, we used bioactive RNTs for siRNA delivery and gene silencing. Fifteen lysine-functionalized twin-G∧C motifs (KnT, = 1 to 15) were synthesized using solid phase peptide synthesis to produce building blocks that self-assembled to produce cationic RNTs under physiological conditions. The intracellular uptake of siRNA delivered by the oligo-L-lysine RNTs was examined and it was found that the complexation of siRNA was affected by the cationic charges from the lysine residues and the length of RNTs formed, with the higher charged KnT RNTs delivering siRNA to the cells at a faster rate. In addition, by protecting siRNA from serum degradation, KnT RNTs were shown to deliver their cargo to the cells effectively the endocytic pathway. A reduction in the expression (∼70%) of the target stat3 protein was observed during gene expression analysis in HCT116 and A549 cell lines.
对遗传性疾病新治疗方法的探索推动了RNA干扰（RNAi）和基因沉默技术的诸多进展。尽管这项技术有解决许多问题的潜力，但其持续应用的关键在于开发有效的递送策略，以降低细胞毒性并提高沉默效率。莲座状纳米管（RNTs）是通过鸟嘌呤 - 胞嘧啶（G∧C）杂交DNA碱基的自组装形成的仿生超分子纳米结构。在此，我们使用生物活性RNTs进行siRNA递送和基因沉默。使用固相肽合成法合成了15个赖氨酸功能化的双G∧C基序（KnT，n = 1至15），以制备在生理条件下自组装产生阳离子RNTs的构建块。研究了由寡聚L - 赖氨酸RNTs递送的siRNA的细胞内摄取情况，发现siRNA的络合受赖氨酸残基的阳离子电荷和形成的RNTs长度的影响，电荷较高的KnT RNTs以更快的速率将siRNA递送至细胞。此外，通过保护siRNA免受血清降解，KnT RNTs被证明能通过内吞途径有效地将其货物递送至细胞。在HCT116和A549细胞系的基因表达分析中，观察到靶标stat3蛋白的表达降低了约70%。
