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用于胶质瘤细胞特异性转染的肽基因递送载体

Peptide Gene Delivery Vectors for Specific Transfection of Glioma Cells.

作者信息

Wang Taoran, Meng Zhao, Kang Ziyao, Ding Guihua, Zhao Baoquan, Han Zhenbin, Zheng Zhibing, Wang Chenhong, Meng Qingbin

机构信息

State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, P. R. China.

Key Laboratory of Natural Resources and Functional Molecules of the Changbai Mountain, Affiliated Ministry of Education, College of Pharmacy, Yanbian University, Yanji, Jilin 133002, China.

出版信息

ACS Biomater Sci Eng. 2020 Dec 14;6(12):6778-6789. doi: 10.1021/acsbiomaterials.0c01336. Epub 2020 Dec 1.

Abstract

Gene therapy offers an alternative approach to malignant glioma; however, glioma cells are difficult to transfect. Peptides, as nonviral vectors, can achieve efficient gene transfection in glioma cells due to their good biocompatibility and easy functionalization. In this article, we reported a series of peptide vectors, which were composed of amphiphilic α-helical segments, cationic cell-penetrating segments, and cysteine and glycine residues. The physicochemical properties of peptide vectors or peptide/pGL3 complexes, including conformation, DNA-loading capacity, size, zeta potential, and morphology, were characterized. Their gene delivery abilities were evaluated in U373, U87, and C6 glioma cell lines and a normal cell line 293 T. Compared with Lipo 2000 and other peptide vectors, the efficiency of P-03 (CLLHHLLHHLLHHGGRKKRRQRRR) to transfect glioma cells was higher. While in 293 T cells, the transfection efficiency of P-03 was much lower than that of Lipo 2000 and another positive control P-07. Furthermore, P-03 could facilitate the pGL3 plasmids crossing a blood-brain barrier model in vitro and achieved the expression of EGFP gene in the brain sites of zebrafish.

摘要

基因治疗为恶性胶质瘤提供了一种替代方法;然而,胶质瘤细胞难以转染。肽作为非病毒载体,由于其良好的生物相容性和易于功能化,能够在胶质瘤细胞中实现高效的基因转染。在本文中,我们报道了一系列肽载体,它们由两亲性α-螺旋片段、阳离子细胞穿透片段以及半胱氨酸和甘氨酸残基组成。对肽载体或肽/pGL3复合物的物理化学性质,包括构象、DNA负载能力、大小、zeta电位和形态进行了表征。在U373、U87和C6胶质瘤细胞系以及正常细胞系293 T中评估了它们的基因递送能力。与Lipofectamine 2000和其他肽载体相比,P-03(CLLHHLLHHLLHHGGRKKRRQRRR)转染胶质瘤细胞的效率更高。而在293 T细胞中,P-03的转染效率远低于Lipofectamine 2000和另一个阳性对照P-07。此外,P-03能够促进pGL3质粒在体外穿过血脑屏障模型,并在斑马鱼的脑部位实现EGFP基因的表达。

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