title: A library of synthetic transcription activator-like effector-activated promoters for coordinated orthogonal gene expression in plants.
abstract: A library of synthetic promoters containing the binding site of a single designer transcription activator-like effector (dTALE) was constructed. The promoters contain a constant sequence, consisting of an 18-base long dTALE-binding site and a TATA box, flanked by degenerate sequences of 49 bases downstream and 19 bases upstream. Forty-three of these promoters were sequenced and tested in transient assays in Nicotiana benthamiana using a GUS reporter gene. The strength of expression of the promoters ranged from around 5% to almost 100% of the viral 35S promoter activity. We then demonstrated the utility of these promoters for metabolic engineering by transiently expressing three genes for the production of a plant diterpenoid in N. benthamiana. The simplicity of the promoter structure shows great promise for the development of genetic circuits, with wide potential applications in plant synthetic biology and metabolic engineering.
authors.affiliation: Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Brückner
authors.affiliation: Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
authors.forename: Petra
authors.initials: P
authors.lastname: Schäfer
authors.affiliation: Icon Genetics GmbH, Weinbergweg 22, 06120, Halle (Saale), Germany.
authors.forename: Ernst
authors.initials: E
authors.lastname: Weber
authors.affiliation: Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
authors.forename: Ramona
authors.initials: R
authors.lastname: Grützner
authors.affiliation: Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
authors.forename: Sylvestre
authors.initials: S
authors.lastname: Marillonnet
authors.affiliation: Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120, Halle (Saale), Germany.
authors.forename: Alain
authors.initials: A
authors.lastname: Tissier
mesh_terms: D018506:Gene Expression Regulation, Plant / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D058615:Synthetic Biology; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
keywords: Nicotiana benthamiana; metabolic engineering; orthogonality; plants; synthetic biology; synthetic promoters; technical advance; terpenoid; transcription activator-like effectors; transient assays
references.citation: Mol Cell. 2014 May 22;54(4):698-710
references.pmid: 24837679
references.citation: Metab Eng. 2014 May;23:145-53
references.pmid: 24704560
references.citation: Nat Rev Microbiol. 2014 Jul;12(7):479-92
references.pmid: 24909109
references.citation: ACS Synth Biol. 2014 Nov 21;3(11):839-43
references.pmid: 24933124
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Science. 2007 Oct 26;318(5850):648-51
references.pmid: 17962565
references.citation: Plant J. 2014 Jun;78(5):753-71
references.pmid: 24602153
references.citation: Bioeng Bugs. 2012 Jan 1;3(1):38-43
references.pmid: 22126803
references.citation: Plant Methods. 2013 Dec 12;9(1):46
references.pmid: 24330621
references.citation: Mol Plant. 2012 Mar;5(2):318-33
references.pmid: 22442388
references.citation: Nat Methods. 2013 Mar;10(3):239-42
references.pmid: 23377379
references.citation: PLoS One. 2011;6(5):e19722
references.pmid: 21625552
references.citation: Plant J. 2009 Sep;59(6):859-71
references.pmid: 19473322
references.citation: EMBO J. 1987 Dec 20;6(13):3901-7
references.pmid: 3327686
references.citation: Curr Opin Microbiol. 2014 Apr;18:83-9
references.pmid: 24637219
references.citation: Nucleic Acids Res. 2013 Jun;41(11):5569-81
references.pmid: 23599004
references.citation: RNA. 2012 Dec;18(12):2299-305
references.pmid: 23105001
references.citation: Science. 2007 Oct 26;318(5850):645-8
references.pmid: 17962564
references.citation: Planta. 2003 Feb;216(4):686-91
references.pmid: 12569411
references.citation: New Phytol. 2013 Jul;199(2):352-66
references.pmid: 23638869
references.citation: PLoS One. 2008;3(11):e3647
references.pmid: 18985154
references.citation: PLoS One. 2012;7(9):e46451
references.pmid: 23029521
references.citation: Cell. 2014 Jun 5;157(6):1262-78
references.pmid: 24906146
references.citation: Nucleic Acids Res. 2012 Aug;40(15):e115
references.pmid: 22730293
references.citation: Plant Mol Biol. 2010 Aug;73(6):673-85
references.pmid: 20495852
references.citation: Science. 2009 Dec 11;326(5959):1509-12
references.pmid: 19933107
references.citation: Annu Rev Phytopathol. 2010;48:419-36
references.pmid: 19400638
references.citation: Nat Biotechnol. 2012 Oct;30(10):1002-6
references.pmid: 22983090
references.citation: Nucleic Acids Res. 1990 Jun 11;18(11):3387-93
references.pmid: 2192362
references.citation: Nat Methods. 2014 May;11(5):508-20
references.pmid: 24781324
references.citation: Plant Mol Biol. 2012 Feb;78(3):311-21
references.pmid: 22167390
references.citation: Gene. 1987;60(2-3):217-25
references.pmid: 2832252
references.citation: Plant Mol Biol. 2010 Jul;73(4-5):363-78
references.pmid: 20354894
references.citation: Biotechnol J. 2013 Jan;8(1):46-58
references.pmid: 22890821
references.citation: PLoS One. 2012;7(3):e33279
references.pmid: 22442681
references.citation: Genetics. 2011 Nov;189(3):705-36
references.pmid: 22084422