Khramtsov Yu V, Lupanova T N, Rosenkranz A A, Georgiev G P, Sobolev A S
Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Moscow State University, Moscow, Russia.
Dokl Biochem Biophys. 2025 Apr;521(1):157-159. doi: 10.1134/S1607672924601136. Epub 2025 Apr 11.
To test new antiviral drugs aimed at degrading the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus, it is desirable to have cells expressing the N-protein, for which it is necessary to find conditions for the maximum achievable efficiency of cell transfection with a plasmid encoding this protein. For transfection, polyplexes were used consisting of a plasmid encoding the N-protein fused with the mRuby3 fluorescent protein and polyethyleneimine (PEI)-polyethylene glycol (PEG)-TAT peptide block copolymers. The dependence of the transfection efficiency of human lung adenocarcinoma A549 cells on the PEG/PEI and N/P ratios (the ratio of nitrogen in PEI to phosphate in DNA) was studied. Significant positive correlations were shown between transfection efficiency determined by flow cytometry, the N/P ratio, and the proportion of polyplexes sized 40-54 nm. The data obtained can serve as a basis for creating an animal model of lung cells transiently expressing the N protein of the SARS-CoV-2 virus.
为了测试旨在降解严重急性呼吸综合征冠状病毒2（SARS-CoV-2）病毒核衣壳蛋白（N蛋白）的新型抗病毒药物，需要有表达N蛋白的细胞，为此有必要找到用编码该蛋白的质粒进行细胞转染时可达到的最大效率的条件。转染时，使用了由编码与mRuby3荧光蛋白融合的N蛋白的质粒和聚乙烯亚胺（PEI）-聚乙二醇（PEG）-TAT肽嵌段共聚物组成的多聚体。研究了人肺腺癌A549细胞的转染效率对PEG/PEI和N/P比率（PEI中氮与DNA中磷酸的比率）的依赖性。通过流式细胞术测定的转染效率、N/P比率与大小为40-54nm的多聚体比例之间显示出显著的正相关。所获得的数据可为创建瞬时表达SARS-CoV-2病毒N蛋白的肺细胞动物模型提供依据。
