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学科主题: 药学
题名:
Single modification at position 14 of siRNA strand abolishes its gene-silencing activity by decreasing both RISC loading and target degradation
作者: Zheng, Jie1; Zhang, Lei1; Zhang, Junbin3; Wang, Xiaoxia1; Ye, Keqiong4; Xi, Zhen3; Du, Quan1,2; Liang, Zicai1
关键词: RNA immunoprecipitation ; AGO ; computational modeling
刊名: FASEB JOURNAL
发表日期: 2013-10-01
DOI: 10.1096/fj.13-228668
卷: 27, 期:10, 页:4017-4026
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biochemistry & Molecular Biology ; Biology ; Cell Biology
研究领域[WOS]: Biochemistry & Molecular Biology ; Life Sciences & Biomedicine - Other Topics ; Cell Biology
关键词[WOS]: CRYSTAL-STRUCTURE ; CHEMICAL-MODIFICATIONS ; MAMMALIAN-CELLS ; COMPLEX ; IMPROVEMENTS ; THERAPEUTICS ; ARGONAUTE2 ; RNAS ; DNA
英文摘要:

Normally siRNA has to be chemically stabilized in therapeutic applications. It is a challenge to obtain optimal stabilizing effects while maintaining full silencing activity due to a lack of understanding of how different chemical modifications would influence the efficacy of siRNA. In the current study, the effect of single 2-sugar modifications was profiled across the length of the siRNA guide strand. This led to the surprising finding that a single 2-OMe modification at position 14 of the siRNA guide strand substantially compromised its gene-silencing activity in a manner that was independent of the nucleotide identity at this site or the sequence context around it. We found that modification at position 14 of the siRNA guide strand reduced its RNA-induced silencing complex (RISC) loading tremendously, whereas the loading of the siRNA sense strand was only marginally affected. When comparing the silencing potency of 14th position-modified siRNA (transfected at 16.7 nM) and native control (transfected at 1 nM) at equivalent Ago2 loading levels, the silencing potency of modified siRNA was much lower, even lower than the level of native siRNA transfected at 0.1 nM. These data indicated that modification at position 14 of the siRNA guide strand abolishes its gene-silencing activity by decreasing both RISC loading and target degradation. Using a computational modeling approach, we demonstrated an intimate interaction between the 14th nucleotide of guide strand and the amino acid Q675 in the AGO protein, which is located in a highly conserved loop of PIWI domain. In addition to gaining insights into siRNA-AGO interactions, this study of structure-activity relationship further established a general principle for siRNA modification in siRNA drug development.Zheng, J., Zhang, L., Zhang, J., Wang, X., Ye, K., Xi, Z., Du, Q., Liang, Z. Single modification at position 14 of siRNA strand abolishes its gene-silencing activity by decreasing both RISC loading and target degradation.

语种: 英语
所属项目编号: 81273422 ; 31221002 ; 2012AA022501 ; 2011CBA01100 ; 2011ZX09102-011-12 ; 2012ZX09102301-006
项目资助者: National Natural Science Foundation of China ; Foundation for Innovative Research Groups of the National Natural Science Foundation of China ; National High-Tech R&amp ; D Program of China ; National Basic Research Program of China ; National Drug Program
WOS记录号: WOS:000329747100016
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内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/57305
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, Inst Mol Med, Sch Pharmaceut Sci, Beijing 100871, Peoples R China
2.Peking Univ, State Key Lab Nat & Biomimet Drugs, Sch Pharmaceut Sci, Beijing 100871, Peoples R China
3.Nankai Univ, State Key Lab Elementoorgan Chem, Tianjin 300071, Peoples R China
4.Natl Inst Biol Sci, Beijing, Peoples R China

Recommended Citation:
Zheng, Jie,Zhang, Lei,Zhang, Junbin,et al. Single modification at position 14 of siRNA strand abolishes its gene-silencing activity by decreasing both RISC loading and target degradation[J]. FASEB JOURNAL,2013,27(10):4017-4026.
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