title: Rapid, modular and reliable construction of complex mammalian gene circuits.
abstract: 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.
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, 40 Ames Street, Cambridge, MA 02142, USA, Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 40 Ames Street, Cambridge, MA 02142, USA and INRIA Paris-Rocquencourt, Le Chesnay, 78153, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Guye
authors.forename: Yinqing
authors.initials: Y
authors.lastname: Li
authors.forename: Liliana
authors.initials: L
authors.lastname: Wroblewska
authors.forename: Xavier
authors.initials: X
authors.lastname: Duportet
authors.forename: Ron
authors.initials: R
authors.lastname: Weiss
mesh_terms: D053263:Gene Regulatory Networks; D005818:Genetic Engineering / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans