合子
微量注射
转基因
胚胎
生物
细胞生物学
胚胎干细胞
绿色荧光蛋白
分子生物学
胶体金
DNA
电穿孔
质粒
后代
基因分型
遗传学
胚胎发生
基因
纳米技术
纳米颗粒
材料科学
基因型
怀孕
作者
Di Zhang,Qi Wang,Xiaocui Cheng,Huanhuan Li,Qiuyue Wang,Fangfang Yu,Wenying Wang,Xin Jing,Qingmei Li,Changfeng Dou,Xiaoqing Wu,Hongli Qian,Wei Fang,Fengrui Wu,Yong Liu,Shuanglin Jiang,Wenyong Li,Rong Wang
出处
期刊:ChemNanoMat
[Wiley]
日期:2020-04-16
卷期号:6 (5): 765-772
被引量:2
标识
DOI:10.1002/cnma.202000007
摘要
Abstract A transgenic system of noninvasive cytosolic transport based on exogenous plasmid DNA (pIRES2−EGFP) loading on gold nanoparticles (AuNPs) coated with polyethyleneimine (PEI) was successfully constructed, which can remove the technical threshold and possible damage of microinjection to oocytes and embryo development. In this study, mouse zygotes were cultured in KSOM medium containing the complex of AuNPs−PEI−DNA (APD) until formation of blastocyts, and then transferred into pseudopregnant female mice for obtaining transgenic offspring. Fluorescence observations confirm the expression of GFP in early embryos; importantly, data from identifying experiments by genotyping, immunofluorescence and DNA sequencing show that the GFP sequence indeed integrates into the chromosome of offspring individuals and is expressed. Furthermore, TUNEL test and analysis on cell differentiation of blastocysts indicate an excellent safety of APD to the development of embryos. All these results suggested that AuNPs, being an alternative technique to traditional microinjection, can efficiently introduce biomolecules into early embryonic cells without the prerequisite of expensive micromanipulators and skilled technicians, which will greatly promote study of life sciences in various aspects.
科研通智能强力驱动
Strongly Powered by AbleSci AI