Polyethylenimine-Mediated CCR5 Gene Knockout Using Transcription Activator-Like Effector Nucleases
文献类型： 外文期刊
作者： Jin, Lian 2 ; Deng, Yan 3 ; He, Nongyue 1 ; Wang, Lijun 4 ; Weng, Mengling 6 ;
作者机构： 1.Southeast Univ, Natl Demonstrat Ctr Expt Biomed Engn Educ, State Key Lab Bioelect, Sch Biol Sci & Med Engn, Nanjing 210096, Jiangsu, Peoples R China
2.Zunyi Med Univ, Bioresource Inst Halth Utilizat, Zunyi 563000, Guizhou, Peoples R China
3.Hunan Univ Technol, Hunan Key Lab Biomed Nanomat & Devices, Econ Forest Cultivat & Utilizat Collaborat Innova, Zhuzhou 412007, Peoples R China
4.Guangxi Univ, Coll Light Ind & Food Engn, Nanning 530004, Peoples R China
5.Guangxi Key Lab Clean Pulp & Paper Making & Pollu, Nanning 530007, Peoples R China
6.Guangxi Sugarcane Res Inst, Nanning 530007, Peoples R China
关键词： CCR5; TALEN; Polyethylenimine; Gene Delivery; HIV
期刊名称：JOURNAL OF BIOMEDICAL NANOTECHNOLOGY （ 影响因子：4.099； 五年影响因子：3.492 ）
ISSN： 1550-7033
年卷期： 2018 年 14 卷 3 期
页码：
收录情况： SCI
摘要： CCR5 acts as one of the key coreceptors for human immunodeficiency virus infection, which leads to acquired immune deficiency syndrome. CCR5 gene knockout comes up as an alternative method for treatment of the disease. Transcription activator-like effector nuclease is a powerful gene editing tool characterized by the ease of design, high rates of cleavage activity, and the accessibility of all ranges of transcription activator-like effector nucleases. In this study, transcription activator-like effector nuclease plasmids specifically targeting to knock out CCR5 gene were designed, constructed and then transfected to Hela and HEK293T cell lines with the assistance of the well-established gene delivery carrier polyethylenimine. The transfection efficiency of polyethylenimine reached 50-60% at the optimized N/P ratio of 6/1. The genomic CCR5 sequencing results suggested an estimated knockout efficiency of around 50% under the optimized condition. Thus, CCR5 gene knockout was achieved in cell lines by the polyethylenimine-mediated transfection of transcription activator-like effector nuclease plasmids of interest. More efforts such as the improvement of transfection efficiency of gene delivery carrier could be made to achieve higher gene knockout efficiency of transcription activator-like effector nucleases.
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