Targeting piggyBac transposon integrations in the human genome.
Targeting piggyBac transposon integrations in the human genome.
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针对人类基因组中的 piggyBac 转座子整合。
DOI：
10.1007/978-1-62703-761-7_9
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Wilson,MatthewH
中科院分区：
文献类型：
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作者：
Galvan,DanielL;Kettlun,ClaudiaS;Wilson,MatthewH
基于DNA的转座子系统提供了一种主动有效地将基因送入人类细胞的技术。与非定向整合相比，一个新兴的领域是操纵这样的系统以实现定点定向整合，非靶向整合发生在天然或未经修饰的转座子系统中。自然激活的芽孢杆菌转座子系统来自昆虫，但已被证明在修改人类细胞基因方面非常有效。最近的努力利用DNA结合域与猪芽孢杆菌转座酶的融合，目标是靶向整合到人类基因组中的特定位置。在本章中，我们描述了设计和表征嵌合猪芽孢杆菌转座酶的方法，包括评估人类基因组中活性和靶向性的实验方法。
DNA based transposon systems offer a technology for active and efficient delivery of genes into human cells. An emerging field is directed at manipulating such systems to achieve site-directed integration as compared to un-targeted integration which occurs with native or unmodified transposon systems. The naturally activepiggyBactransposon system is derived from insects but has been shown to be very efficient in gene-modifying human cells. Recent efforts have utilized the fusion of DNA binding domains to thepiggyBactransposase enzyme with the goal of targeting integration to specific locations in the human genome. In this chapter, we describe methodology for engineering and characterizing chimericpiggyBactransposase enzymes, including experimental approaches for evaluating activity and targeting specificity in the human genome.