新型整合素ανβ3靶向siRNA载体的构建及生物学评价

Xueqi Chen, Meng Liu, Rongfu Wang, P. Yan, Chun-li Zhang, Chao Ma, L. Yin
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引用次数: 6

摘要

(1)背景:基于RNA干扰(RNAi)的基因治疗的巨大潜力是以小干扰RNA (sirna)有效递送到靶组织和细胞为前提的。因此,我们旨在开发和检测一种新的整合素αvβ3特异性递送载体,用于靶向转染siRNA到恶性肿瘤细胞;(2)方法:采用精氨酸-甘氨酸-天冬氨酸基序(RGD)作为组织靶点特异性识别整合素αvβ3。为了使siRNA结合,在环RGD二聚体的羧基端添加精氨酸残基(9R)合成了一个嵌合肽,命名为c(RGD)2-9R。通过流式细胞术、共聚焦显微镜和Western blot对9R肽转移siRNA的效率进行体外生物学评价;(3)结果:琼脂糖凝胶电泳证实了c(RGD)2-9R与siRNA的最佳摩尔比为10:1。流式细胞术和共聚焦显微镜结果证实,转染c(RGD)2-9R作为siRNA递送载体的转染率明显高于裸siRNA组。Western blot结果表明,这些9R肽能够在体外将siRNA转导到HepG2细胞中,从而实现有效的基因沉默;(4)结论:c(RGD)2-9R嵌合肽可作为siRNA的有效递送载体,有望成为基于rnai的基因治疗的新策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Construction and Biological Evaluation of a Novel Integrin ανβ3-Specific Carrier for Targeted siRNA Delivery In Vitro
(1) Background: The great potential of RNA interference (RNAi)-based gene therapy is premised on the effective delivery of small interfering RNAs (siRNAs) to target tissues and cells. Hence, we aimed at developing and examining a novel integrin αvβ3-specific delivery carrier for targeted transfection of siRNA to malignant tumor cells; (2) Methods: Arginine-glycine-aspartate motif (RGD) was adopted as a tissue target for specific recognition of integrin αvβ3. To enable siRNA binding, a chimeric peptide was synthesized by adding nonamer arginine residues (9R) at the carboxy terminus of cyclic-RGD dimer, designated as c(RGD)2-9R. The efficiency of 9R peptide transferring siRNA was biologically evaluated in vitro by flow cytometry, confocal microscopy, and Western blot; (3) Results: An optimal 10:1 molar ratio of c(RGD)2-9R to siRNA was confirmed by the electrophoresis on agarose gels. Both the flow cytometry and confocal microscopy results testified that transfection of c(RGD)2-9R as an siRNA delivery carrier was obviously higher than the naked-siRNA group. The results of Western blot demonstrated that these 9R peptides were able to transduce siRNA to HepG2 cells in vitro, resulting in efficient gene silencing; and (4) Conclusion: The chimeric peptide of c(RGD)2-9R can be developed as an effective siRNA delivery carrier and shows potential as a new strategy for RNAi-based gene therapy.
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