聚乙烯亚胺
转染
HEK 293细胞
质粒
绿色荧光蛋白
Cas9
基因传递
细胞毒性
分子生物学
生物物理学
清脆的
化学
细胞生物学
组合化学
生物
体外
生物化学
DNA
基因
作者
Akbar Hassanzadeh,Amir Hossein Vahabi,Seyyed Emad Hooshmand,Elaheh Hosseini,Behjat Kheiri Yeghaneh Azar,Jafar Kiani,Sara Saeedi,Ali Shahbazi,Arnab Rudra,Michael R. Hamblin,Karimi
出处
期刊:Nanotechnology
[IOP Publishing]
日期:2022-07-26
卷期号:33 (44): 445101-445101
标识
DOI:10.1088/1361-6528/ac842d
摘要
Abstract Despite the development of many novel carriers for the delivery of various types of genetic material, the lack of a delivery system with high efficiency and low cytotoxicity is a major bottleneck. Herein, low molecular weight polyethylenimine (PEI 1.8k ) was functionalized with saponin residues using phenylboronic acid (PBA) as an ATP-responsive cross-linker, and a fluorinated side chain to construct PEI-PBA-SAP-F polycation as a highly efficient delivery vector. This vehicle could transfect small plasmid DNA (∼3 kb) with outstanding efficiency into various cells, including HEK 293T, NIH3T3, A549, PC12, MCF7 and HT-29, as well as robust transfection of a large plasmid (∼9 kb) into HEK 293T cells. The carrier indicated good transfection efficacy even at high concentration of serum and low doses of plasmid. The use of green fluorescent protein (GFP) knock-out analysis demonstrated transfection of different types of CRISPR/Cas9 complexes (Cas9/sgRNA ribonucleoproteins RNP, plasmid encoding Cas9 plus sgRNA targeting GFP, Cas9 expression plasmid plus in vitro -prepared sgRNA). In summary, we report an effective PEI-PBA-SAP-F gene carrier with the appropriate lipophilic/cationic balance for biomedical applications.
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