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Silencing effect of shRNA expression vectors with stem length of 21, 27 or 29 bp

文献类型： 外文期刊

作者： Liu, Zhonghua 2 ; Xiao, Hongwei 1 ; Zheng, Xinmin 1 ; Qiao, Xianfeng 1 ; Wang, Huayan 2 ;

作者机构： 1.Hubei Acad Agr Sci, Inst Anim & Vet Sci, Wuhan 430064, Peoples R China

2.NW A&F Univ, Shaanxi Stem Cell Engn & Technol Res Ctr, Shaanxi Key Lab Mol Biol Agr, Coll Vet Med, Yangling 712100, Peoples R China

3.Hubei Key Lab Anim Embryo & Mol Breeding, Wuhan 430064, Peoples R China

关键词： Gene silenging;shRNA;enhanced green fluorescent protein;mouse embryonic fibroblast

期刊名称：AFRICAN JOURNAL OF BIOTECHNOLOGY （ 影响因子：0.573； 五年影响因子：0.794 ）

ISSN： 1684-5315

年卷期： 2011 年 10 卷 7 期

页码：

收录情况： SCI

摘要： In this study, shRNA vectors having different stem length were constructed and their silencing effect was tested in mouse embryonic fibroblast and in vivo. Interfering RNAs were designed with stems of 21, 27, and 29 bp. The enhanced green fluorescent protein gene was used as target gene. The synthesized single strands were annealed and cloned into psiSTRIKE. Then, the recombinant plasmids were transfected into mouse embryonic fibroblast with lipofection and injected into leg muscle of mouse. The mRNA expression level of the green fluorescent protein gene was checked by real-time quantitative polymerase chain reaction (RT-PCR). The silencing effect of the 29 bp shRNA vector was stronger than that of the 21 and 27 bp in cell, but there was no significant difference among them when injected in muscle. The findings in this study are of interest for selecting the best hairpins for mouse individuals.

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[2]不同长度茎部shRNA表达载体构建与鉴定. 刘中华,乔宪凤,肖红卫,刘西梅,王华岩,郑新民. 2010

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