Institute of Microbiology, Czech Academy of Sciences, Prague, Czech Republic.
Center of Molecular Structure, Institute of Biotechnology, Czech Academy of Sciences, Vesec, Czech Republic.
Sci Rep. 2023 Sep 6;13(1):14669. doi: 10.1038/s41598-023-41567-7.
In this study, gold nanoparticles produced by eukaryotic cell waste (AuNP), were analyzed as a transfection tool. AuNP were produced by Fusarium oxysporum and analyzed by spectrophotometry, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). Fourier transform infrared spectroscopy (FTIR) and dynamic light scattering (DLS) were used before and after conjugation with different nucleic acid (NA) types. Graphite furnace atomic absorption spectroscopy (GF-AAS) was used to determine the AuNP concentration. Conjugation was detected by electrophoresis. Confocal microscopy and quantitative real-time PCR (qPCR) were used to assess transfection. TEM, SEM, and EDS showed 25 nm AuNP with round shape. The amount of AuNP was 3.75 ± 0.2 µg/µL and FTIR proved conjugation of all NA types to AuNP. All the samples had a negative charge of - 36 to - 46 mV. Confocal microscopy confirmed internalization of the ssRNA-AuNP into eukaryotic cells and qPCR confirmed release and activity of carried RNA.
在这项研究中，我们分析了真核细胞废物产生的金纳米颗粒（AuNP）作为转染工具。AuNP 是由尖孢镰刀菌产生的，并通过分光光度法、透射电子显微镜（TEM）、扫描电子显微镜（SEM）和能谱分析（EDS）进行了分析。在与不同核酸（NA）类型结合前后，使用傅里叶变换红外光谱（FTIR）和动态光散射（DLS）进行了分析。使用石墨炉原子吸收光谱（GF-AAS）测定 AuNP 浓度。通过电泳检测结合。使用共聚焦显微镜和实时荧光定量 PCR（qPCR）评估转染。TEM、SEM 和 EDS 显示出 25nm 的圆形 AuNP。AuNP 的数量为 3.75 ± 0.2μg/μL，FTIR 证明了所有 NA 类型与 AuNP 的结合。所有样品的负电荷为-36 至-46 mV。共聚焦显微镜证实了 ssRNA-AuNP 被真核细胞内化，qPCR 证实了携带的 RNA 的释放和活性。
