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肽衍生各向异性金纳米颗粒作为高效 siRNA 载体。

Peptide generated anisotropic gold nanoparticles as efficient siRNA vectors.

机构信息

School of Biotechnology, Kalinga Institute of Industrial Technology Deemed to be University, Campus 11, Patia, Bhubaneswar, Odisha 751024, India.

Conversion & Catalysis Division, CSIR-Indian Institute of Petroleum, Dehradun 248005, India.

出版信息

Int J Pharm. 2019 May 30;563:198-207. doi: 10.1016/j.ijpharm.2019.04.007. Epub 2019 Apr 3.

Abstract

Based on the cell penetrating ability of tryptophan-containing peptides, eight linear hexapeptides have been designed, synthesized and explored their efficiency toward the synthesis of gold nanoparticles under sunlight. The peptide generated gold nanoparticles (LP-GNPs) have been characterized by UV-visible spectroscopy, Transmission Electron Microscopy (TEM) and Dynamic Light Scattering (DLS) techniques. The binding ability of LP-GNPs toward siRNA, evaluated by gel electrophoresis indicates that sequence-selective-GNPs without any surface modifications exhibit strong affinity toward negatively charged biomolecules. Cellular uptake studies suggest that LP-GNPs exhibit significant uptake of fluorescence-labeled siRNA inside the cells as evidenced from Fluorescence Microscopy. In vitro gene silencing efficiency using newly generated GNPs revealed that above mentioned LP-GNPs efficiently down-regulate the level of GAPGH gene in colon cancer cells. Comparative gene silencing efficiency results indicate that anisotropic LP7-GNPs exhibit comparable efficacy to other existing carrier systems, such as Lipofectamine 2000 in presence of serum, mimicking in-vivo system. In conclusion, our results demonstrate that peptide-GNPs based delivery system for siRNA emerges to be effective to deliver RNAi therapeutics, uncovering new avenue in oncotherapy.

摘要

基于色氨酸肽的细胞穿透能力,设计、合成了 8 条线性六肽,并探索了它们在阳光照射下合成金纳米粒子的效率。肽生成的金纳米粒子(LP-GNPs)通过紫外-可见光谱、透射电子显微镜(TEM)和动态光散射(DLS)技术进行了表征。通过凝胶电泳评估 LP-GNPs 对 siRNA 的结合能力表明,未经任何表面修饰的序列选择性-GNPs 对带负电荷的生物分子具有很强的亲和力。细胞摄取研究表明,LP-GNPs 在内皮细胞内表现出对荧光标记 siRNA 的显著摄取,荧光显微镜证实了这一点。使用新生成的 GNPs 进行体外基因沉默效率研究表明,上述 LP-GNPs 可有效下调结肠癌细胞中 GAPGH 基因的水平。比较基因沉默效率的结果表明,各向异性的 LP7-GNPs 在存在血清的情况下与其他现有载体系统(如 Lipofectamine 2000)具有相当的功效,模拟了体内系统。总之,我们的结果表明,基于肽-GNPs 的 siRNA 传递系统在传递 RNAi 治疗方面是有效的,为肿瘤治疗开辟了新途径。

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