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.

中文翻译：

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AuNPs-PEI对转基因小鼠的DNA无损递送至合子

成功构建了基于外源质粒DNA（pIRES2-EGFP）的无创胞质转运转基因系统，该质粒负载在涂有聚乙烯亚胺（PEI）的金纳米颗粒（AuNPs）上，该技术可以消除技术门槛以及显微注射对卵母细胞和胚胎发育的可能损害。在这项研究中，小鼠合子在含有AuNPs-PEI-DNA（APD）复合物的KSOM培养基中培养直到形成囊胚，然后转移到假孕雌性小鼠中以获得转基因后代。荧光观察证实了GFP在早期胚胎中的表达。重要的是，通过基因分型，免疫荧光和DNA测序鉴定实验得到的数据表明，GFP序列确实整合到了后代个体的染色体中并得以表达。此外，TUNEL测试和对胚泡细胞分化的分析表明，APD对胚胎发育具有极好的安全性。所有这些结果表明，AuNPs是传统显微注射的一种替代技术，可以将生物分子有效地引入到早期胚胎细胞中，而无需昂贵的显微操纵器和熟练的技术人员，这将极大地促进生命科学研究的各个方面。