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学科主题: 药学
题名:
D-Isonucleotide (isoNA) incorporation around cleavage site of passenger strand promotes the vibration of Ago2-PAZ domain and enhances in vitro potency of siRNA
作者: Huang, Ye1,5,6; Tian, Miao1; Zhang, Yichao2; Sheng, Gang3; Chen, Zhuo1; Ma, Yuan1; Chen, Yue1; Peng, Yihong4; Zhao, Yi-Lei2; Wang, Yanli3; Zhang, Lihe1; Yang, Zhenjun1
刊名: ORGANIC & BIOMOLECULAR CHEMISTRY
发表日期: 2015
DOI: 10.1039/c5ob01119a
卷: 13, 期:44, 页:10825-10833
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Chemistry, Organic
研究领域[WOS]: Chemistry
关键词[WOS]: SILENCING ACTIVITY ; RNA INTERFERENCE ; RISC ; STABILITY ; EXPRESSION ; COMPLEXES ; PROTEIN ; CELLS
英文摘要:

It has been demonstrated that passenger strand cleavage is important for the activation of RNA-induced silencing complex (RISC), which is a crucial step for siRNA-mediated gene silencing. Herein, we report that isonucleotide (isoNA) modification around the cleavage site of the passenger strand would affect the in vitro potency of modified siRNAs by altering the motion pattern of the Ago2-PAZ domain. According to western blotting, q-PCR and antiviral test results, we proved that D-isonucleotide (isoNA) modification at the position 8 of the passenger strand (siMek1-S08D), which is adjacent to the cleavage site, markedly improved the in vitro potency of the modified siRNA, whereas siRNAs with D-isoNA incorporation at position 9 (siMek1-S09D) or L-isoNA incorporation at positions 8 and 9 (siMek1-S08L, siMek1-S09L) displayed lower activity compared to native siRNA. Kinetics evaluation of passenger strand cleavage induced by T. thermophilus Ago (Tt-Ago) showed that D-isoNA modification at position 8 of the passenger strand had no significant influence on the cleavage rate, but L-isoNA modification at position 8 slowed the cleavage rate markedly. Moreover, the results of molecular dynamics simulations showed that D-isoNA modification at position 8 affected the open-close motion of the PAZ domain in the Ago/siRNA complex, which may promote the loading of RISC and release of a passenger strand cleavage product, and consequently accelerate the activation of RISC and enhance silencing activity. However, D-isoNA modification at position 9 or L-isoNA modification at position 8 or 9 exerted opposite influences on the motion of the Ago-PAZ domain.

语种: 英语
所属项目编号: 2012AA022501 ; 2012CB720604 ; 2013CB966800 ; 2013CB966802 ; 20932001 ; 21377085
项目资助者: Ministry of Science and Technology of China ; National Natural Science Foundation of China
WOS记录号: WOS:000364077400006
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/62414
Appears in Collections:北京大学药学院_期刊论文

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作者单位: 1.Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China
2.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai 200030, Peoples R China
3.Chinese Acad Sci, Inst Biophys, Lab Noncoding RNA, Beijing 100080, Peoples R China
4.Peking Univ, Sch Basic Med Sci, Dept Microbiol, Beijing 100871, Peoples R China
5.Peking Union Med Coll, Inst Mat Med, Dept New Drug Res & Dev, Beijing 100021, Peoples R China
6.Chinese Acad Med Sci, Beijing 100730, Peoples R China

Recommended Citation:
Huang, Ye,Tian, Miao,Zhang, Yichao,et al. D-Isonucleotide (isoNA) incorporation around cleavage site of passenger strand promotes the vibration of Ago2-PAZ domain and enhances in vitro potency of siRNA[J]. ORGANIC & BIOMOLECULAR CHEMISTRY,2015,13(44):10825-10833.
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