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Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial

机译:果蝇中P元素介导的转基因效率提高:显微注射与纳米材料复合的DNA

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摘要

The efficiency of genetic transformation technology to generate stable transgenics depends upon the successful delivery of plasmid DNA in embryonic cells. The available gene vectors facilitate efficient plasmid DNA delivery to the cellular milieu but are exposed to nuclease degradation. Recent in vitro studies suggest encapsulation of plasmid DNA with nanomaterial(s) for better protection against nucleases. Therefore, in this study, we tested if complexing of free plasmid DNA with linear polyethylenimine (LPEI, 25 kDa) based nanoparticle (LPN) enhances the efficiency of transformation (transgenesis) by using Drosophila based germ-line transformation technology. Here, we show that the LPN-DNA complex not only enhances the efficiency of this transgenic technology at a DNA concentration of 0.04 μg/μl but also reduces the DNA quantity required to generate transgenics by ten folds. This approach has potential applications for other types of transgenesis and nucleic acid injection methods in Drosophila as well as other popular genetic model systems.
机译:遗传转化技术产生稳定转基因的效率取决于质粒DNA在胚胎细胞中的成功传递。可用的基因载体有助于将质粒DNA有效地递送至细胞环境,但是暴露于核酸酶降解。最近的体外研究表明,用纳米材料包裹质粒DNA可以更好地保护核酸酶。因此,在这项研究中,我们测试了使用基于果蝇的种系转化技术,将游离质粒DNA与基于线性聚乙烯亚胺(LPEI,25 kDa)的纳米颗粒(LPN)的复合是否增强转化(转基因)的效率。在这里,我们显示LPN-DNA复合物不仅在0.04μg/μl的DNA浓度下提高了这种转基因技术的效率,而且将生成转基因所需的DNA量减少了十倍。这种方法对于果蝇以及其他流行的遗传模型系统中的其他类型的转基因和核酸注射方法具有潜在的应用。

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