A red light-controlled synthetic gene expression switch for plant systems.

On command control of gene expression in time and space is required for the comprehensive analysis of key plant cellular processes. Even though some chemical inducible systems showing satisfactory induction features have been developed, they are inherently limited in terms of spatiotemporal resolution and may be associated with toxic effects. We describe here the first synthetic light-inducible system for the targeted control of gene expression in plants. For this purpose, we applied an interdisciplinary synthetic biology approach comprising mammalian and plant cell systems to customize and optimize a split transcription factor based on the plant photoreceptor phytochrome B and one of its interacting factors (PIF6). Implementation of the system in transient assays in tobacco protoplasts resulted in strong (95-fold) induction in red light (660 nm) and could be instantaneously returned to the OFF state by subsequent illumination with far-red light (740 nm). Capitalizing on this toggle switch-like characteristic, we demonstrate that the system can be kept in the OFF state in the presence of 740 nm-supplemented white light, opening up perspectives for future application of the system in whole plants. Finally we demonstrate the system's applicability in basic research, by the light-controlled tuning of auxin signalling networks in N. tabacum protoplasts, as well as its biotechnological potential for the chemical-inducer free production of therapeutic proteins in the moss P. patens.

[1]  H G Kang,et al.  A glucocorticoid-inducible transcription system causes severe growth defects in Arabidopsis and induces defense-related genes. , 1999, The Plant journal : for cell and molecular biology.

[2]  Christopher A. Voigt,et al.  Synthetic biology: Engineering Escherichia coli to see light , 2005, Nature.

[3]  Differential Top10 promoter regulation by six tetracycline analogues in plant cells. , 2002, Journal of experimental botany.

[4]  Björn H. Junker,et al.  Synthetic gene networks in plant systems. , 2012, Methods in molecular biology.

[5]  Rainer Fischer,et al.  Plant-based production of biopharmaceuticals. , 2004, Current opinion in plant biology.

[6]  Malra C. Treece.  An A is an A is an A is an E. , 1976 .

[7]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[8]  M. Gore,et al.  Chemical-Inducible, Ecdysone Receptor-Based Gene Expression System for Plants , 2003, Transgenic Research.

[9]  Matias D. Zurbriggen,et al.  Synthesis of phycocyanobilin in mammalian cells. , 2013, Chemical communications.

[10]  R. Reski,et al.  A gene responsible for prolyl-hydroxylation of moss-produced recombinant human erythropoietin , 2013, Scientific Reports.

[11]  Paul Christou,et al.  Genetic modification: The production of recombinant pharmaceutical proteins in plants , 2003, Nature Reviews Genetics.

[12]  T. Halazonetis,et al.  Ultraviolet-B-mediated induction of protein–protein interactions in mammalian cells , 2013, Nature Communications.

[13]  V. Mett,et al.  Copper-controllable gene expression system for whole plants. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Reski,et al.  Optimisation of a bioreactor culture of the moss Physcomitrella patens for mass production of protoplasts , 2002 .

[15]  Ahmad S. Khalil,et al.  Synthetic biology: applications come of age , 2010, Nature Reviews Genetics.

[16]  M. Fussenegger,et al.  Functional cross-kingdom conservation of mammalian and moss (Physcomitrella patens) transcription, translation and secretion machineries. , 2009, Plant biotechnology journal.

[17]  A. McDonald,et al.  A glucocorticoid-inducible gene expression system can cause growth defects in tobacco , 2007, Planta.

[18]  R. Rosenfeld.  Nature , 2009, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[19]  R. Reski,et al.  Plant nuclear gene knockout reveals a role in plastid division for the homolog of the bacterial cell division protein FtsZ, an ancestral tubulin. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Christopher A. Voigt,et al.  Multichromatic control of gene expression in Escherichia coli. , 2011, Journal of molecular biology.

[21]  R. Reski,et al.  Enhanced recovery of a secreted recombinant human growth factor using stabilizing additives and by co-expression of human serum albumin in the moss Physcomitrella patens. , 2005, Plant biotechnology journal.

[22]  M. Rodriguez-Franco,et al.  High‐level expression of secreted complex glycosylated recombinant human erythropoietin in the Physcomitrella Δ‐fuc‐t Δ‐xyl‐t mutant , 2007 .

[23]  Randall J. Platt,et al.  Optical Control of Mammalian Endogenous Transcription and Epigenetic States , 2013, Nature.

[24]  J. Sheen,et al.  Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis , 2007, Nature Protocols.

[25]  A. Hartmann,et al.  Production of biologically active recombinant human factor H in Physcomitrella. , 2011, Plant biotechnology journal.

[26]  S. Goff,et al.  Regulation of gene expression by small molecules in rice. , 2001, Novartis Foundation symposium.

[27]  Yi-shin Su,et al.  Phytochrome structure and signaling mechanisms. , 2006, Annual review of plant biology.

[28]  C. Fankhauser,et al.  Phosphorylation of Phytochrome B Inhibits Light-Induced Signaling via Accelerated Dark Reversion in Arabidopsis[W][OA] , 2013, Plant Cell.

[29]  N. Chua,et al.  Technical advance: An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. , 2000, The Plant journal : for cell and molecular biology.

[30]  M. Fussenegger,et al.  Novel pristinamycin-responsive expression systems for plant cells. , 2001, Biotechnology and bioengineering.

[31]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[32]  R. Reski,et al.  Moss-based production of asialo-erythropoietin devoid of Lewis A and other plant-typical carbohydrate determinants. , 2012, Plant biotechnology journal.

[33]  E. Huq,et al.  A light-switchable gene promoter system , 2002, Nature Biotechnology.

[34]  Andreas Möglich,et al.  From dusk till dawn: one-plasmid systems for light-regulated gene expression. , 2012, Journal of molecular biology.

[35]  Anna Whyatt,et al.  Notes and references , 1984, International Journal of Legal Information : Official Publication.

[36]  N. Chua,et al.  A glucocorticoid-mediated transcriptional induction system in transgenic plants. , 1997, The Plant journal : for cell and molecular biology.

[37]  C. Fankhauser,et al.  Phytochrome-mediated light signalling in Arabidopsis. , 2004, Current opinion in plant biology.

[38]  L. H. Grimme,et al.  Tetrapyrrole biosynthesis in algae and higher plants: a discussion of the importance of the 5-aminolaevulinate synthase and the dioxovalerate transaminase pathways in the biosynthesis of chlorophyll. , 1978, The International journal of biochemistry.

[39]  G. Corrado,et al.  Inducible gene expression systems and plant biotechnology. , 2009, Biotechnology advances.

[40]  Gary Walsh,et al.  Biopharmaceutical benchmarks 2010 , 2010, Nature Biotechnology.

[41]  Christian S. Hardtke,et al.  Hormone Signalling Crosstalk in Plant Growth Regulation , 2011, Current Biology.

[42]  Yi Yang,et al.  Spatiotemporal control of gene expression by a light-switchable transgene system , 2012, Nature Methods.

[43]  J. Botto,et al.  The plant cell , 2007, Plant Molecular Biology Reporter.

[44]  Wilfried Weber,et al.  Optogenetic tools for mammalian systems. , 2013, Molecular bioSystems.

[45]  Matias D. Zurbriggen,et al.  Plant and bacterial systems biology as platform for plant synthetic bio(techno)logy. , 2012, Journal of biotechnology.

[46]  Klaus Palme,et al.  A quantitative ratiometric sensor for time-resolved analysis of auxin dynamics , 2013, Scientific Reports.

[47]  A. Cigan,et al.  A Chimeric Ecdysone Receptor Facilitates Methoxyfenozide-Dependent Restoration of Male Fertility in ms45 Maize , 2002, Transgenic Research.

[48]  M. Padidam,et al.  Chemically regulated gene expression in plants. , 2003, Current opinion in plant biology.

[49]  C. Stemmer,et al.  In vivo glyco‐engineered antibody with improved lytic potential produced by an innovative non‐mammalian expression system , 2007, Biotechnology journal.

[50]  R. Reski,et al.  Glycoprotein production in moss bioreactors , 2011, Plant Cell Reports.

[51]  Jens Timmer,et al.  Multi-chromatic control of mammalian gene expression and signaling , 2013, Nucleic acids research.

[52]  C. Glover,et al.  Gene expression profiling for hematopoietic cell culture , 2006 .

[53]  B. Felenbok.  The ethanol utilization regulon of Aspergillus nidulans: the alcA-alcR system as a tool for the expression of recombinant proteins. , 1991, Journal of biotechnology.

[54]  F. Nagy,et al.  Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter , 1985, Nature.

[55]  Martin Fussenegger,et al.  SAMY, a novel mammalian reporter gene derived from Bacillus stearothermophilus α-amylase , 2002 .

[56]  Stefan Kepinski,et al.  Defining auxin response contexts in plant development. , 2010, Current opinion in plant biology.

[57]  C. D. de Oliveira,et al.  A combinatorial TIR1/AFB-Aux/IAA co-receptor system for differential sensing of auxin , 2012, Nature chemical biology.

[58]  Matias D. Zurbriggen,et al.  Novel perspectives for the engineering of abiotic stress tolerance in plants. , 2014, Current opinion in biotechnology.

[59]  R. Tanaka,et al.  Tetrapyrrole biosynthesis in higher plants. , 2007, Annual review of plant biology.

[60]  M. Gossen,et al.  A chimeric transactivator allows tetracycline-responsive gene expression in whole plants. , 1994, The Plant journal : for cell and molecular biology.

[61]  H. Koop,et al.  Integration of foreign sequences into the tobacco plastome via polyethylene glycol-mediated protoplast transformation , 2004, Planta.

[62]  N. Ferrara,et al.  The biology of vascular endothelial growth factor. , 1997, Endocrine reviews.

[63]  Effect of Ploidy Level on Growth, Differentiation, and Morphology in Physcomitrella patens , 2005 .

[64]  Miss A.O. Penney.  (b) , 1974, The New Yale Book of Quotations.

[65]  W. Marsden.  I and J , 2012 .

[66]  Jens Timmer,et al.  A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells , 2013, Nucleic acids research.

[67]  M. Fussenegger,et al.  Macrolide-based transgene control in mammalian cells and mice , 2002, Nature Biotechnology.

[68]  M. Ehlers,et al.  Rapid blue light induction of protein interactions in living cells , 2010, Nature Methods.