高内涵筛选用于评价新型siRNA运载试剂的细胞群体生物学活性

doi:10.5246/jcps.2013.06.068

journal1

高内涵筛选用于评价新型siRNA运载试剂的细胞群体生物学活性

李雅婷, 郑宜, 潘德林, 徐波, 武芸^*, 杨振军, 张礼和

北京大学医学部天然药物及仿生药物国家重点实验室, 北京 100191

收稿日期:2013-05-29 修回日期:2013-06-17 出版日期:2013-11-15 发布日期:2014-01-22
通讯作者: 武芸*

Development of a high-content screening method to evaluate the cellular population-based biological activity of new siRNA delivering reagent

Yating Li, Yi Zheng, Delin Pan, Bo Xu, Yun Wu^*, Zhenjun Yang, Lihe Zhang

State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2013-05-29 Revised:2013-06-17 Online:2013-11-15 Published:2014-01-22
Contact: Yun Wu*

摘要/Abstract

摘要： 修饰siRNAs和新型siRNA运载系统对细胞的作用通常通过PCR、Western Blotting和荧光显微镜来进行细胞整体特征的研究。本文报道了基于高内涵、用于siRNA治疗的一种新型运载系统的新策略(CLD), 该策略从细胞整体和细胞群体水平综合考虑siRNA的转染效率及对蛋白表达水平的影响。我们证明CLD能达到更佳的细胞摄取, 使siBraf达到更好的抗肿瘤活性。该方法具有高效、准确和适用于多个贴壁细胞系的优点, 能简化siRNA的研究过程

关键词: 高内涵筛选, siRNA运载系统, 基因敲减, 细胞群体, ERK信号通路

Abstract: Normally, cellular responses to modified siRNAs or new siRNA delivery systems have been studied in group cell behavior by PCR, western blotting and fluorescence microscopy. In this study, we present a novel high-content screening (HCS) strategy to evaluate a novel delivery system (named CLD) of siRNA therapeutics, with which both the content of intracellular siRNAs and changes in protein expressing levels have been quantified in group cells and cellular population. We also observed that with the better cell uptake, CLD provided siRNA therapeutics (siBraf) better antitumor capability. This novel strategy was proved to be with efficiency, accuracy and high competency to adherent cell lines, thus making siRNA research more simplified

Key words: High-content screening, siRNA delivery system, Gene knock-down, Cellular population, ERK signaling pathway

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Supporting:

Foundation items: Ministry of Science and Technology of China (Grant No. 2012CB720604) and NSFC (Grant No. 20932001). #br# ^*Corresponding author. Tel.: 86-10-82805023

李雅婷, 郑宜, 潘德林, 徐波, 武芸^*, 杨振军, 张礼和. 高内涵筛选用于评价新型siRNA运载试剂的细胞群体生物学活性[J]. journal1, DOI: 10.5246/jcps.2013.06.068.

Yating Li, Yi Zheng, Delin Pan, Bo Xu, Yun Wu^*, Zhenjun Yang, Lihe Zhang . Development of a high-content screening method to evaluate the cellular population-based biological activity of new siRNA delivering reagent[J]. Journal of Chinese Pharmaceutical Sciences, DOI: 10.5246/jcps.2013.06.068.

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高内涵筛选用于评价新型siRNA运载试剂的细胞群体生物学活性

Development of a high-content screening method to evaluate the cellular population-based biological activity of new siRNA delivering reagent

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