编码SARS-CoV-2病毒n蛋白质粒转染A549细胞效率的优化

IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yu. V. Khramtsov, T. N. Lupanova, A. A. Rosenkranz,  G. P. Georgiev,  A. S. Sobolev
{"title":"编码SARS-CoV-2病毒n蛋白质粒转染A549细胞效率的优化","authors":"Yu. V. Khramtsov,&nbsp;T. N. Lupanova,&nbsp;A. A. Rosenkranz,&nbsp; G. P. Georgiev,&nbsp; A. S. Sobolev","doi":"10.1134/S1607672924601136","DOIUrl":null,"url":null,"abstract":"<p>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.</p>","PeriodicalId":529,"journal":{"name":"Doklady Biochemistry and Biophysics","volume":"521 1","pages":"157 - 159"},"PeriodicalIF":0.8000,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Optimization of A549 Cell Transfection Efficiency with a Plasmid Encoding the N-Protein of the SARS-CoV-2 Virus\",\"authors\":\"Yu. V. Khramtsov,&nbsp;T. N. Lupanova,&nbsp;A. A. Rosenkranz,&nbsp; G. P. Georgiev,&nbsp; A. S. Sobolev\",\"doi\":\"10.1134/S1607672924601136\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>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.</p>\",\"PeriodicalId\":529,\"journal\":{\"name\":\"Doklady Biochemistry and Biophysics\",\"volume\":\"521 1\",\"pages\":\"157 - 159\"},\"PeriodicalIF\":0.8000,\"publicationDate\":\"2025-04-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Doklady Biochemistry and Biophysics\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1134/S1607672924601136\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Doklady Biochemistry and Biophysics","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1134/S1607672924601136","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

为了测试旨在降解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,为此需要找到用编码该蛋白的质粒转染细胞的最高效率的条件。转染时,采用由编码n蛋白与mRuby3荧光蛋白融合的质粒和聚乙烯亚胺(PEI)-聚乙二醇(PEG)-TAT肽段共聚物组成的多聚体。研究了人肺腺癌A549细胞转染效率与PEG/PEI和N/P (PEI中氮与DNA中磷酸盐的比值)的关系。流式细胞术检测的转染效率、N/P比值与40-54 nm大小的复合物比例呈显著正相关。获得的数据可作为建立瞬时表达SARS-CoV-2病毒N蛋白的肺细胞动物模型的基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Optimization of A549 Cell Transfection Efficiency with a Plasmid Encoding the N-Protein of the SARS-CoV-2 Virus

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.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Doklady Biochemistry and Biophysics
Doklady Biochemistry and Biophysics 生物-生化与分子生物学
CiteScore
1.60
自引率
12.50%
发文量
68
审稿时长
6-12 weeks
期刊介绍: Doklady Biochemistry and Biophysics is a journal consisting of English translations of articles published in Russian in biochemistry and biophysics sections of the Russian-language journal Doklady Akademii Nauk. The journal''s goal is to publish the most significant new research in biochemistry and biophysics carried out in Russia today or in collaboration with Russian authors. The journal accepts only articles in the Russian language that are submitted or recommended by acting Russian or foreign members of the Russian Academy of Sciences. The journal does not accept direct submissions in English.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信