{"title":"Optimized PEI-based Transfection Method for Transient Transfection and Lentiviral Production.","authors":"Shaozhe Yang, Xiaoling Zhou, Rongxiang Li, Xiuhong Fu, Pingnan Sun","doi":"10.1002/cpch.25","DOIUrl":null,"url":null,"abstract":"<p><p>Polyethyleneimine (PEI), a cationic polymer vehicle, forms a complex with DNA which then can carry anionic nucleic acids into eukaryotic cells. PEI-based transfection is widely used for transient transfection of plasmid DNA. The efficiency of PEI-based transfection is affected by numerous factors, including the way the PEI/DNA complex is prepared, the ratio of PEI to DNA, the concentration of DNA, the storage conditions of PEI solutions, and more. Considering the major influencing factors, PEI-based transfection has been optimized to improve its efficiency, reproducibility, and consistency. This protocol outlines the steps for ordinary transient transfection and lentiviral production using PEI. © 2017 by John Wiley & Sons, Inc.</p>","PeriodicalId":38051,"journal":{"name":"Current protocols in chemical biology","volume":"9 3","pages":"147-157"},"PeriodicalIF":0.0000,"publicationDate":"2017-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpch.25","citationCount":"28","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in chemical biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1002/cpch.25","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 28
Abstract
Polyethyleneimine (PEI), a cationic polymer vehicle, forms a complex with DNA which then can carry anionic nucleic acids into eukaryotic cells. PEI-based transfection is widely used for transient transfection of plasmid DNA. The efficiency of PEI-based transfection is affected by numerous factors, including the way the PEI/DNA complex is prepared, the ratio of PEI to DNA, the concentration of DNA, the storage conditions of PEI solutions, and more. Considering the major influencing factors, PEI-based transfection has been optimized to improve its efficiency, reproducibility, and consistency. This protocol outlines the steps for ordinary transient transfection and lentiviral production using PEI. © 2017 by John Wiley & Sons, Inc.
基于pei的瞬时转染和慢病毒生产方法优化。
聚乙烯亚胺(PEI)是一种阳离子聚合物载体,与DNA形成复合物,然后将阴离子核酸携带到真核细胞中。基于pei的转染被广泛用于质粒DNA的瞬时转染。PEI转染的效率受到许多因素的影响,包括PEI/DNA复合物的制备方式、PEI与DNA的比例、DNA的浓度、PEI溶液的储存条件等等。考虑到主要的影响因素,我们对pei转染进行了优化,以提高其效率、重复性和一致性。本协议概述了使用PEI进行普通瞬时转染和慢病毒生产的步骤。©2017 by John Wiley & Sons, Inc。
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