title: Transferring a synthetic gene circuit from yeast to mammalian cells.
abstract: The emerging field of synthetic biology builds gene circuits for scientific, industrial and therapeutic needs. Adaptability of synthetic gene circuits across different organisms could enable a synthetic biology pipeline, where circuits are designed in silico, characterized in microbes and reimplemented in mammalian settings for practical usage. However, the processes affecting gene circuit adaptability have not been systematically investigated. Here we construct a mammalian version of a negative feedback-based 'linearizer' gene circuit previously developed in yeast. The first naïve mammalian prototype was non-functional, but a computational model suggested that we could recover function by improving gene expression and protein localization. After rationally developing and combining new parts as the model suggested, we regained function and could tune target gene expression in human cells linearly and precisely as in yeast. The steps we have taken should be generally relevant for transferring any gene circuit from yeast into mammalian cells.
authors: Department of Systems Biology-Unit 950, The University of Texas MD Anderson Cancer Center, 7435 Fannin Street, Houston, Texas 77054, USA.
authors: Dmitry
authors: D
authors: Nevozhay
authors: Tomasz
authors: T
authors: Zal
authors: Gábor
authors: G
authors: Balázsi
mesh_terms: D000818:Animals; D001483:Base Sequence; D003062:Codon / Q000235:genetics; D015870:Gene Expression; D053263:Gene Regulatory Networks; D005800:Genes, Fungal / Q000235:genetics; D017930:Genes, Reporter; D005813:Genes, Synthetic; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007438:Introns / Q000235:genetics; D061986:MCF-7 Cells; D008322:Mammals / Q000378:metabolism*; D008958:Models, Molecular; D016254:Mutagenesis, Insertional / Q000235:genetics; D019913:Nuclear Localization Signals / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D003062:Codon; D019913:Nuclear Localization Signals; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
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