{"title":"Transfection by Electroporation","authors":"Huntington Potter, Richard Heller","doi":"10.1002/cpmb.48","DOIUrl":null,"url":null,"abstract":"<p>Electroporation—the use of high-voltage electric shocks to introduce DNA into cells—can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit describes electroporation of mammalian cells, including ES cells for the preparation of knock-out, knock-in, and transgenic mice. Protocols are described for the use of electroporation in vivo to perform gene therapy for cancer therapy and DNA vaccination. Also described are modifications for preparation and transfection of plant protoplasts. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10734,"journal":{"name":"Current Protocols in Molecular Biology","volume":"121 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmb.48","citationCount":"53","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmb.48","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 53
Abstract
Electroporation—the use of high-voltage electric shocks to introduce DNA into cells—can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit describes electroporation of mammalian cells, including ES cells for the preparation of knock-out, knock-in, and transgenic mice. Protocols are described for the use of electroporation in vivo to perform gene therapy for cancer therapy and DNA vaccination. Also described are modifications for preparation and transfection of plant protoplasts. © 2018 by John Wiley & Sons, Inc.
电穿孔转染
电穿孔——使用高压电击将DNA导入细胞——可以用于大多数细胞类型,产生高频率的稳定转化和瞬时基因表达,而且,因为它需要更少的步骤,比其他技术更容易。本单元描述了哺乳动物细胞的电穿孔,包括用于制备敲除、敲入和转基因小鼠的胚胎干细胞。描述了在体内使用电穿孔进行癌症治疗和DNA疫苗接种的基因治疗的方案。还描述了制备和转染植物原生质体的修饰。©2018 by John Wiley &儿子,Inc。
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