title: A transcription activator-like effector (TALE) induction system mediated by proteolysis.
abstract: Simple and predictable trans-acting regulatory tools are needed in the fields of synthetic biology and metabolic engineering to build complex genetic circuits and optimize the levels of native and heterologous gene products. Transcription activator-like effectors (TALEs) are bacterial virulence factors that have recently gained traction in biotechnology applications owing to their customizable DNA-binding specificity. In this work we expanded the versatility of these transcription factors to create an inducible TALE system by inserting tobacco-etch virus (TEV) protease recognition sites into the TALE backbone. The resulting engineered TALEs maintain transcriptional repression of their target genes in Escherichia coli, but are degraded after induction of the TEV protease, thereby promoting expression of the previously repressed target gene of interest. This TALE-TEV technology enables both repression and induction of plasmid or chromosomal target genes in a manner analogous to traditional repressor proteins but with the added flexibility of being operator-agnostic.
authors: University of Wisconsin-Madison, Department of Chemical and Biological Engineering, Madison, Wisconsin, USA.
authors: Matthew F
authors: MF
authors: Copeland
authors: University of Wisconsin-Madison, Department of Chemical and Biological Engineering, Madison, Wisconsin, USA.
authors: Mark C
authors: MC
authors: Politz
authors: University of Wisconsin-Madison, Department of Chemical and Biological Engineering, Madison, Wisconsin, USA.
authors: Charles B
authors: CB
authors: Johnson
authors: University of Wisconsin-Madison, Department of Chemical and Biological Engineering, Madison, Wisconsin, USA.
authors: Andrew L
authors: AL
authors: Markley
authors: University of Wisconsin-Madison, Department of Chemical and Biological Engineering, Madison, Wisconsin, USA.
authors: Brian F
authors: http://orcid.org/0000-0002-9232-9959
authors: BF
authors: Pfleger
mesh_terms: D010450:Endopeptidases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D059748:Proteolysis; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D058615:Synthetic Biology / Q000379:methods*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D012097:Repressor Proteins; D014157:Transcription Factors; D037521:Virulence Factors; D010450:Endopeptidases; C099488:TEV protease