title: Modular construction of mammalian gene circuits using TALE transcriptional repressors.
abstract: An important goal of synthetic biology is the rational design and predictable implementation of synthetic gene circuits using standardized and interchangeable parts. However, engineering of complex circuits in mammalian cells is currently limited by the availability of well-characterized and orthogonal transcriptional repressors. Here, we introduce a library of 26 reversible transcription activator-like effector repressors (TALERs) that bind newly designed hybrid promoters and exert transcriptional repression through steric hindrance of key transcriptional initiation elements. We demonstrate that using the input-output transfer curves of our TALERs enables accurate prediction of the behavior of modularly assembled TALER cascade and switch circuits. We also show that TALER switches using feedback regulation exhibit improved accuracy for microRNA-based HeLa cancer cell classification versus HEK293 cells. Our TALER library is a valuable toolkit for modular engineering of synthetic circuits, enabling programmable manipulation of mammalian cells and helping elucidate design principles of coupled transcriptional and microRNA-mediated post-transcriptional regulation.
authors: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 40 Ames St, Cambridge MA 02142, USA.
authors: Yinqing
authors: Y
authors: Li
authors: Bioinformatics Division/Center for Synthetic & Systems Biology, Tsinghua National Laboratory for Information Science and Technology, Tsinghua University, Beijing, 100084, China.
authors: Yun
authors: Y
authors: Jiang
authors: Bioinformatics Division/Center for Synthetic & Systems Biology, Tsinghua National Laboratory for Information Science and Technology, Tsinghua University, Beijing, 100084, China.
authors: He
authors: H
authors: Chen
authors: Bioinformatics Division/Center for Synthetic & Systems Biology, Tsinghua National Laboratory for Information Science and Technology, Tsinghua University, Beijing, 100084, China.|MOE Key Laboratory of Bioinformatics; Department of Automation, Tsinghua University, Beijing 100084, China.
authors: Weixi
authors: W
authors: Liao
authors: Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, 935 Jiaoling Road, Kunming, Yunnan, 650118.
authors: Zhihua
authors: Z
authors: Li
authors: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, 40 Ames St, Cambridge MA 02142, USA.|Department of Biological Engineering, Massachusetts Institute of Technology, 40 Ames St, Cambridge MA 02142, USA.
authors: Ron
authors: R
authors: Weiss
authors: Bioinformatics Division/Center for Synthetic & Systems Biology, Tsinghua National Laboratory for Information Science and Technology, Tsinghua University, Beijing, 100084, China.|MOE Key Laboratory of Bioinformatics; Department of Automation, Tsinghua University, Beijing 100084, China.
authors: Zhen
authors: Z
authors: Xie
mesh_terms: D000818:Animals; D001483:Base Sequence; D053263:Gene Regulatory Networks / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008322:Mammals; D008969:Molecular Sequence Data; D015202:Protein Engineering / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics*; D054852:Small Molecule Libraries; D015534:Trans-Activators / Q000235:genetics*; D061785:Transcription Initiation, Genetic
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D054852:Small Molecule Libraries; D015534:Trans-Activators
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