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  • Synthetic Biology and Assembly Cloning  (2)
  • Belief, J61 - Geographic Labor Mobility  (1)
  • Targeted gene modification  (1)
  • Oxford University Press  (4)
  • Institute of Physics (IOP)
  • 1
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    A platform for rapid prototyping of synthetic gene networks in mammalian cells (2014)
    Oxford University Press
    Details
    Publication Date: 2014-11-28
    Description: Mammalian synthetic biology may provide novel therapeutic strategies, help decipher new paths for drug discovery and facilitate synthesis of valuable molecules. Yet, our capacity to genetically program cells is currently hampered by the lack of efficient approaches to streamline the design, construction and screening of synthetic gene networks. To address this problem, here we present a framework for modular and combinatorial assembly of functional (multi)gene expression vectors and their efficient and specific targeted integration into a well-defined chromosomal context in mammalian cells. We demonstrate the potential of this framework by assembling and integrating different functional mammalian regulatory networks including the largest gene circuit built and chromosomally integrated to date (6 transcription units, 27kb) encoding an inducible memory device. Using a library of 18 different circuits as a proof of concept, we also demonstrate that our method enables one-pot/single-flask chromosomal integration and screening of circuit libraries. This rapid and powerful prototyping platform is well suited for comparative studies of genetic regulatory elements, genes and multi-gene circuits as well as facile development of libraries of isogenic engineered cell lines.
    Keywords: Synthetic Biology and Assembly Cloning
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 2
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    Rapid, modular and reliable construction of complex mammalian gene circuits (2013)
    Oxford University Press
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    Publication Date: 2013-09-06
    Description: We developed a framework for quick and reliable construction of complex gene circuits for genetically engineering mammalian cells. Our hierarchical framework is based on a novel nucleotide addressing system for defining the position of each part in an overall circuit. With this framework, we demonstrate construction of synthetic gene circuits of up to 64 kb in size comprising 11 transcription units and 33 basic parts. We show robust gene expression control of multiple transcription units by small molecule inducers in human cells with transient transfection and stable chromosomal integration of these circuits. This framework enables development of complex gene circuits for engineering mammalian cells with unprecedented speed, reliability and scalability and should have broad applicability in a variety of areas including mammalian cell fermentation, cell fate reprogramming and cell-based assays.
    Keywords: Synthetic Biology and Assembly Cloning
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 3
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    Highly efficient homology-driven genome editing in human T cells by combining zinc-finger nuclease mRNA and AAV6 donor delivery (2016)
    Oxford University Press
    Details
    Publication Date: 2016-02-20
    Description: The adoptive transfer of engineered T cells for the treatment of cancer, autoimmunity, and infectious disease is a rapidly growing field that has shown great promise in recent clinical trials. Nuclease-driven genome editing provides a method in which to precisely target genetic changes to further enhance T cell function in vivo. We describe the development of a highly efficient method to genome edit both primary human CD8 and CD4 T cells by homology-directed repair at a pre-defined site of the genome. Two different homology donor templates were evaluated, representing both minor gene editing events (restriction site insertion) to mimic gene correction, or the more significant insertion of a larger gene cassette. By combining zinc finger nuclease mRNA delivery with AAV6 delivery of a homologous donor we could gene correct 41% of CCR5 or 55% of PPP1R12C (AAVS1) alleles in CD8 + T cells and gene targeting of a GFP transgene cassette in 〉40% of CD8 + and CD4 + T cells at both the CCR5 and AAVS1 safe harbor locus, potentially providing a robust genome editing tool for T cell-based immunotherapy.
    Keywords: Targeted gene modification
    Print ISSN: 0305-1048
    Electronic ISSN: 1362-4962
    Topics: Biology
    Published by Oxford University Press
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  • 4
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    Global cluster networks--foreign direct investment flows from Canada to China (2013)
    Oxford University Press
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    Publication Date: 2013-12-19
    Description: Using a network perspective of multinational firms, this article develops conceptions of global cluster networks and global city-region networks that are based on foreign direct investment (FDI) activities. The article first formulates a global cluster-network hypothesis suggesting that multinational cluster firms are more likely to set up new foreign affiliates in other, similarly specialized clusters to keep up with global industry dynamics. Conversely, it is suggested that non-cluster firms are more likely to avoid cluster destinations in their FDIs. Second, it is hypothesized that cluster networks generate connections between city-regions in different countries that are horizontal and vertical in character and thus shape global city-region networks. To test these hypotheses, the spatial patterns of 299 FDI cases from Canada to China between 2006 and 2010 are investigated, generally supporting the hypotheses developed.
    Keywords: D83 - Search ; Learning ; Information and Knowledge ; Communication ; Belief, J61 - Geographic Labor Mobility ; Immigrant Workers, R12 - Size and Spatial Distributions of Regional Economic Activity
    Print ISSN: 1468-2702
    Electronic ISSN: 1468-2710
    Topics: Geography , Economics
    Published by Oxford University Press
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