The effect of serum in culture on RNAi efficacy through modulation of polyplexes size.
The effect of serum in culture on RNAi efficacy through modulation of polyplexes size.
复制标题
培养物中的血清通过调节复合物大小对 RNAi 功效的影响。
DOI：
10.1016/j.biomaterials.2013.09.102
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发表时间：
2014
期刊：
影响因子：
14
通讯作者：
Xu, Haiyan
中科院分区：
文献类型：
--
作者：
Liu, Jian;Tabata, Yasuhiko;Meng, Jie;Xu, Haiyan
培养液中的血清是影响非病毒载体RNAi效率的关键因素之一。然而，血清在siRNA传递中的作用机制仍不清楚。在这项工作中，我们以一种阳离子聚合物精胺修饰的普鲁兰多糖(称为普鲁兰-精胺，Ps)作为siRNA载体模型，研究血清对siRNA传递关键步骤的影响，包括Ps和siRNA多聚体(Ps-siRNA)的形成、细胞摄取、溶酶体逃逸和细胞毒性。结果表明，较低的血清浓度(1.25%和2.5%)会产生较大的Ps-siRNA颗粒，而较高的血清浓度(10%-40%)则会产生较小的Ps-siRNA颗粒。与小颗粒相比，大颗粒更有效地启动了siRNA的内化。包埋的PS-siRNA颗粒主要分布在溶酶体中。大颗粒比小颗粒表现出更强的破坏溶酶体稳定的能力，因为大颗粒含有更多的胺，从而引发更强的质子海绵效应，从而导致更有效的siRNA溶酶体逃逸。尽管RNAi效率较低，但在高血清培养条件下，小颗粒的Ps-siRNA产生的细胞毒性要低得多。这些发现解释了为什么血清显著影响RNAi，并提出了一种通过调节血清浓度和增强溶酶体失稳来提高RNAi效率和安全性的策略。
Serum in the culture medium is one crucial factor that compromises RNAi efficiency of non-viral vectors. However, mechanistic roles of serum in siRNA delivery remain unknown. In this work, we took one cationic polymer, pullulan chemically modified by spermine (termed as pullulan-spermine, Ps), as a siRNA carrier model to investigate the effects of serum on key steps in siRNA delivery including formation of Ps and siRNA polyplexes (Ps-siRNA), cellular uptake, lysosomal escape, and cytotoxicity. We demonstrate that low serum concentration (1.25% and 2.5%) in culture medium results in large particles of Ps-siRNA, while high serum concentration (10%–40%) leads to small particles of Ps-siRNA. The larger particles initiated the internalization of siRNA more effectively in comparison to the smaller ones. The engulfed Ps-siRNA particles mainly locate in lysosomes. The large particles exhibited stronger abilities of destabilizing lysosomes than that of the small particles as large Ps-siRNA particles contain more amines and subsequently elicit a stronger proton sponge effect which results in more effective lysosomal escape of siRNA. Despite the lower RNAi efficiency, the small particle of Ps-siRNA in the high serum medium generates much lower cytotoxicity. These findings explain why serum significantly affects RNAi and also propose a strategy for improving RNAi efficiency and safety by modulating serum concentration and enhancing lysosomal destabilization.
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影响因子：
10.8
作者：
Park, Ji Sun;Han, Tae Hee;Cho, Yong Woo
通讯作者：
Cho, Yong Woo
影响因子：
10.8
作者：
Jo, Jun-ichiro;Kai, Tomonori;Tabata, Yasuhiko
通讯作者：
Tabata, Yasuhiko
影响因子：
10.8
作者：
Kanatani, Isao;Ikai, Tomonori;Tabata, Yasuhiko
通讯作者：
Tabata, Yasuhiko
影响因子：
5.1
作者：
Ogris, M;Brunner, S;Wagner, E
通讯作者：
Wagner, E
DOI：
10.1016/j.jconrel.2012.05.022
发表时间：
2012-08
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Chinmay Pangarkar;A. Dinh;S. Mitragotri
通讯作者：
Chinmay Pangarkar;A. Dinh;S. Mitragotri