Rapid, modular and reliable construction of complex mammalian gene circuits | Zendy

Patrick Guye | Zendy; Yinqing Li | Zendy; Liliana Wróblewska | Zendy; Xavier Duportet | Zendy; Ron Weiss | Zendy

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Rapid, modular and reliable construction of complex mammalian gene circuits

Author(s) -

Patrick Guye,

Yinqing Li,

Liliana Wróblewska,

Xavier Duportet,

Ron Weiss

Publication year - 2013

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkt605

Subject(s) - biology , reprogramming , synthetic biology , computational biology , modular design , gene regulatory network , gene , transcription factor , regulation of gene expression , transfection , gene expression , microbiology and biotechnology , genetics , computer science , operating system

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.

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