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Updated: Jun 19, 2026

Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells
Published on: December 4, 2007
Positive selection of DNA-protein interactions in mammalian cells through phenotypic coupling with retrovirus
Ulrich Tschulena1, Kenneth R Peterson, Beatriz Gonzalez
1The Skaggs Institute for Chemical Biology and Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, USA.
Abstract:
Through the shuffling of predefined modular zinc finger domains with predictable target site recognition in vitro, we have generated a large repertoire of artificial transcription factors with five zinc finger domains (TF(ZF)s). Here we report an effective strategy for the selection of ATF libraries by coupling expression of transcriptional activators of the promoter of interest to the enhanced production of retroviral vector particles transferring the TF(ZF) encoding gene. Using this strategy, we successfully selected specific TF(ZF)s that upregulate the expression of the gamma-globin promoter. Selected transcription factors induced the expression of gamma-globin when coupled to an activation domain and reduced expression when linked to a repression domain. This new retroviral approach might be used to select other TF(ZF)s but might also be generalized for the selection of other protein and small-molecule interactions.
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