Medimmune Limited

Adam J Brown

David James

Diane Hatton

Suzanne J Gibson

David C James

As demonstrated herein, when composite transcription factor binding sites do not function synergistically, mammalian promoters can be constructed according to simple design rules. Host-cell transcriptional machinery components were analyzed in silico to identify transcription factors with desired expression dynamics. Cognate binding sites were then comprehensively tested in homotypic and heterotypic architectures to assess modularity and determine the transcriptional activity exhibited by a single copy of each site. When elements were specifically selected to prevent combinatorial interactions, heterotypic promoter activities could be accurately modeled simply as a function of constituent binding site copy numbers. As binding site order, spacing, and orientation had minimal effect on promoter activity, blocks could be optimally combined and arranged in silico according to context-specific design-criteria. To demonstrate this, CHO cell promoters were created de-novo that exhibited designed activity levels and long-term expression stability in vitro.

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Sheffield, United Kingdom

Adam J Brown