title: Libraries of Synthetic TALE-Activated Promoters: Methods and Applications.
abstract: The discovery of proteins with programmable DNA-binding specificities triggered a whole array of applications in synthetic biology, including genome editing, regulation of transcription, and epigenetic modifications. Among those, transcription activator-like effectors (TALEs) due to their natural function as transcription regulators, are especially well-suited for the development of orthogonal systems for the control of gene expression. We describe here the construction and testing of libraries of synthetic TALE-activated promoters which are under the control of a single TALE with a given DNA-binding specificity. These libraries consist of a fixed DNA-binding element for the TALE, a TATA box, and variable sequences of 19 bases upstream and 43 bases downstream of the DNA-binding element. These libraries were cloned using a Golden Gate cloning strategy making them usable as standard parts in a modular cloning system. The broad range of promoter activities detected and the versatility of these promoter libraries make them valuable tools for applications in the fine-tuning of expression in metabolic engineering projects or in the design and implementation of regulatory circuits.
authors.affiliation: Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany.
authors.forename: T
authors.initials: T
authors.lastname: Schreiber
authors.affiliation: Leibniz Institute of Plant Biochemistry, Halle (Saale), Germany. Electronic address: alain.tissier@ipb-halle.de.
authors.forename: A
authors.initials: A
authors.lastname: Tissier
mesh_terms: D060054:Agrobacterium / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D010944:Plants / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D058615:Synthetic Biology / Q000379:methods; D000071336:Transcription Activator-Like Effectors / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D000071336:Transcription Activator-Like Effectors; D004247:DNA
keywords: Metabolic engineering; Orthogonal regulation of gene expression; Plants; Regulatory circuits; Synthetic promoter libraries; Transcription activator-like effectors