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{"title":"Genetic Engineering of Human Pluripotent Stem Cells Using PiggyBac Transposon System","authors":"Mi Ae Park, Ho Sun Jung, Igor Slukvin","doi":"10.1002/cpsc.63","DOIUrl":null,"url":null,"abstract":"<p>Human pluripotent stem cells (hPSCs) emerged as an important tool to investigate human development and disease. These studies often require genetically engineering hPSCs to stably express a transgene, which remains functional in various hPSC progeny. PiggyBac transposon is a highly effective and technically simple vector system with large cargo space available for permanent gene delivery. This unit describes the use of PiggyBac transposons to genetically engineer hPSCs to introduce conditionally expressed transgene or reporter to effectively monitor gene expression during differentiation. Both methods enable robust generation of stable hPSC lines within 1 month. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":53703,"journal":{"name":"Current Protocols in Stem Cell Biology","volume":"47 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpsc.63","citationCount":"14","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Stem Cell Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpsc.63","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: 14
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Abstract
Human pluripotent stem cells (hPSCs) emerged as an important tool to investigate human development and disease. These studies often require genetically engineering hPSCs to stably express a transgene, which remains functional in various hPSC progeny. PiggyBac transposon is a highly effective and technically simple vector system with large cargo space available for permanent gene delivery. This unit describes the use of PiggyBac transposons to genetically engineer hPSCs to introduce conditionally expressed transgene or reporter to effectively monitor gene expression during differentiation. Both methods enable robust generation of stable hPSC lines within 1 month. © 2018 by John Wiley & Sons, Inc.
利用PiggyBac转座子系统进行人类多能干细胞的基因工程
人类多能干细胞(hPSCs)成为研究人类发育和疾病的重要工具。这些研究通常需要对人造血干细胞进行基因工程,以稳定地表达一种转基因,这种转基因在各种人造血干细胞后代中保持功能。PiggyBac转座子是一种高效且技术简单的载体系统,具有较大的载货空间,可用于永久基因传递。本单元描述了使用PiggyBac转座子对hPSCs进行基因工程,以引入有条件表达的转基因或报告基因,以有效监测分化过程中的基因表达。两种方法都能在1个月内生成稳定的hPSC细胞系。©2018 by John Wiley &儿子,Inc。
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