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

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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.