Cys2-His2型锌指蛋白安全运输近红外荧光蛋白进入哺乳动物细胞
The Security of Cys2-His2 Zinc Finger Protein Transports near Infrared Fluorescent Protein into Mammalian Cells
庞晓楠 1李世汉 1贾子瑄 1佟玉玺 1刘佳 2陈喜文 3陈德富1
作者信息
- 1. 南开大学生命科学学院,遗传学和细胞生物学系,天津 300071
- 2. 上海科技大学免疫化学研究所,上海 201210
- 3. 南开大学生命科学学院,生物化学与分子生物学系,天津 300071
- 折叠
摘要
细胞穿膜肽(cell-penetrating peptides,CPPs)是一类能够运输核酸、蛋白及化学药物进入细胞内的短肽.Cys2-His2型锌指蛋白(zinc finger protein,ZFP)具有穿膜肽功能,但其转运的目标分子在细胞内的分布及其对细胞活力的影响尚未见报道.将ZFP与近红外荧光蛋白iRFP682融合表达并评估其通过哺乳动物细胞膜的特性.结果发现,ZFP介导的iRFP682能够进入哺乳动物细胞并定位于细胞核.随着ZFP个数的增加,细胞内的荧光信号强度随之增强,且ZFP作为穿膜肽未对哺乳动物细胞活力产生影响.ZFP介导的细胞穿膜过程与细胞种类不存在相关性,并有望应用于活体成像研究.
Abstract
Cell-penetrating peptides(CPPs)are a kind of short peptides that can transport nucleic acids,proteins and chemical drugs into mammalian cells.Cys2-His2 zinc finger protein(ZFP)could function as a CPP,but the distribution of target cargo in cells and the effects on cell viability have not been reported.The fusion protein of ZFP and near-infrared fluorescent protein iRFP682 was expressed in E.coli cell.Then the transduction efficiency of iRFP682 mediated by ZFP into mammalian cells was evaluated.After purified and transduced into mammalian cells,ZFPn-iRFP682 was located to the nuclear.Comparison of the transduction efficiency revealed that the fluorescence intensity was increased with the increase of ZFP number.ZFPn-iRFP682 did not affect viability of the receipt cell.Therefore,ZFP-mediated transmembrane processes are not correlated with cell types and could be expected to use in vivo imaging studies.
关键词
Cys2-His2型锌指蛋白/细胞穿膜肽/穿膜效率/iRFP682/哺乳动物细胞Key words
Cys2-His2 zinc finger protein/cell-penetrating peptides/transduction efficiency/iRFP682/mammalian cell引用本文复制引用
出版年
2024
NETL
NSTL
万方数据