POWRE: Reusable Selection Cassettes for Gene Integration or Disruption in Mammalian Systems

这项提议的目标是开发一种用于哺乳动物细胞的基因整合盒，它消除了DNA工程中常见的抗性选择问题。PI之前的所有工作都是在酵母中进行的，这一提议代表着向完全不同的哺乳动物系统迈出的重要一步。一个可重复使用的选择盒将用于基因插入，之后将被移除并用于后续整合。这项技术的使用将在两种细胞系--CHO细胞和一种人肺泡上皮细胞系中进行评估。

The goal of this proposal is to develop a gene integration cassette for mammalian cells which eliminates the resistance selection problem common in DNA engineering efforts. All of the PI's previous work has been in yeast, and this proposal represents a significant move into mammalian systems which are quite different. A reusable selection cassette will be used for the gene insertion after which it will be removed and used for subsequent integrations. The use of this technique will be evaluated in two cell lines, CHO cells and a human alveolar epithelial cell line.