Split-TALE: A TALE-Based Two-Component System for Synthetic Biology Applications in Planta

Schreiber, Tom; Prange, Anja; Hoppe, Tina; Tissier, Alain

doi:10.1104/pp.18.01218

Cited by 16 publications

(16 citation statements)

References 60 publications

(110 reference statements)

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“…A. tumefaciens strains containing the respective constructs were cultured according to Schreiber et al (2019) and diluted to an OD 600 of 0.4. The constructs were mixed in a 1:1 ratio before injection into mature leaves of 4-week-old N. benthamiana plants using a needleless syringe.…”

Section: Cloning Of the Tps10-encoding Gene And Creation Of Fusion Comentioning

confidence: 99%

Medicago TERPENE SYNTHASE 10 Is Involved in Defense Against an Oomycete Root Pathogen

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Section: Cloning Of the Tps10-encoding Gene And Creation Of Fusion Comentioning

confidence: 99%

Medicago TERPENE SYNTHASE 10 Is Involved in Defense Against an Oomycete Root Pathogen

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“…All other JAZ proteins did not show an interaction with SlMYB21 in the BiFC assays (not shown). The interaction of SlMYB21 with JAZ9 was further validated by the split transcription activator-like effector(TALE) system (Schreiber et al, 2019). The fusion of SlMYB21 to the activation domain (AD) and of JAZ9 to the BD of TALE did not result in induction of the reporter construct (Supplemental Figure 4).…”

Section: Jasmonate-regulated Slmyb21 Encodes a Flower-specific Transcmentioning

confidence: 97%

“…BiFC binary vectors were achieved using 2in1 vectors according to Grefen and Blatt, 2012. For splitTALE assay (Schreiber et al, 2019), the CDS of MYB21 with or without AD (C-terminal deletion of 25 aa) and JAZ9 were cloned into level 1 vectors using Golden Gate method (Werner et al, 2012), resulting in a fusion either to the TALE BD (N-terminal; Act2p:TALE(BD)-MYB21) or to AD (C-terminal; 35S:JAZ9-TALE[AD]; Schreiber and Tissier, 2016). For splitTALE assays, constructs were transformed into Agrobacterium tumefaciens strain GV3101 and used for transient expression in Nicotiana benthamiana leaves.…”

Section: Cloning Of Myb21 and Jaz-encoding Genes For Interaction Studiesmentioning

confidence: 99%

“…Overnight cultured Agrobacteria transformed with the respective plasmids were resuspended in infiltration media consisting of 10 mM MES, 10 mM MgCl 2 , and 100 mM acetosyringone to OD 600 = 0.4. Agrobacteria containing the splitTALE constructs and the reporter construct 4xSTAP1:GUS (Schreiber and Tissier, 2016;Schreiber et al, 2019) were mixed in ratio 1:1:1 and used to transiently transform N. benthamiana leaves through syringemediated infiltration (Sparkes et al, 2006). At 3 d after inoculation, leaf material was harvested, frozen in liquid nitrogen, homogenized, and mixed with extraction buffer containing 50 mM Na 2 HPO 4 (pH 7), 10 mM EDTA, 0.1% (w/v) SDS, 0.1% (v/v) Triton X-100, and 10 mM b-mercaptoethanol.…”

Section: Splittale Assays In N Benthamianamentioning

confidence: 99%

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Tomato MYB21 Acts in Ovules to Mediate Jasmonate-Regulated Fertility

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The function of the plant hormone jasmonic acid (JA) in the development of tomato (Solanum lycopersicum) flowers was analyzed with a mutant defective in JA perception (jasmonate-insensitive1-1, jai1-1). In contrast with Arabidopsis (Arabidopsis thaliana) JA-insensitive plants, which are male sterile, the tomato jai1-1 mutant is female sterile, with major defects in female development. To identify putative JA-dependent regulatory components, we performed transcriptomics on ovules from flowers at three developmental stages from wild type and jai1-1 mutants. One of the strongly downregulated genes in jai1-1 encodes the MYB transcription factor SlMYB21. Its Arabidopsis ortholog plays a crucial role in JA-regulated stamen development. SlMYB21 was shown here to exhibit transcription factor activity in yeast, to interact with SlJAZ9 in yeast and in planta, and to complement Arabidopsis myb21-5. To analyze SlMYB21 function, we generated clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR associated protein 9 (Cas9) mutants and identified a mutant by Targeting Induced Local Lesions in Genomes (TILLING). These mutants showed female sterility, corroborating a function of MYB21 in tomato ovule development. Transcriptomics analysis of wild type, jai1-1, and myb21-2 carpels revealed processes that might be controlled by SlMYB21. The data suggest positive regulation of JA biosynthesis by SlMYB21, but negative regulation of auxin and gibberellins. The results demonstrate that SlMYB21 mediates at least partially the action of JA and might control the flower-to-fruit transition.

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“…nonnative) synthetic sensing and regulatory circuits and their application to plant development. The review by Andres et al (2019) introduces and illustrates the key concepts of synthetic switches, logic gates, and regulatory circuits, which leads neatly to the Research Articles by Iacopino et al (2019), on the design and testing of a synthetic oxygen sensor for plants based on bioengineered versions of a mammalian oxygen sensor, and by Schreiber et al (2019), on design and optimization of a two-component AND-gate system for synthetic circuits in plants based on bacterial transcription activator-like effectors. Wright and Nemhauser (2019) review how SynBio can help uncover the parts and logic mediating plant development, leading to predictive models that specify the molecular circuitry needed to change cell state.…”

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Focus Issue Editorial: Synthetic Biology

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Synthetic biology (SynBio) is a conceptual and operational revolution (Church et al., 2014) that's coming soon to a branch of plant science near you, if it's not there already (Liu and Stewart, 2015). The Synthetic Biology Focus Issue sets out to spread this disruptive news. SynBio is a transformative combination of DNA technology, engineering principles, and computational tools that makes it possible to design new life processes and to repurpose existing natural ones for useful purposes (Purnick and Weiss, 2009). SynBio will profoundly impact and empower how plant science is done and how plant science is used to sustainably solve global problems. SynBio is already creating new jobs and is likely to keep doing this for decades (Delebecque and Philp, 2015). SynBio is powerful for conceptual and operational reasons. The enormous conceptual power of SynBio is to open access to the vast "design space" that plants, and nature in general, have not explored (Bhatia et al., 2017). By discovering and deploying useful design space that evolution has "missed," SynBio enables plants: to make familiar compounds by new pathways, and make new-to-nature compounds; to supply genes needed to make high-value compounds in microorganisms; to manipulate familiar morphological structures, and build totally new-to-nature ones; to respond in new ways to old stimuli, and sense and respond to totally new stimuli; and to be managed based on weather forecasts and other predictions that plants cannot make themselves. The transformative operational power of SynBio lies in its drive to industrialize biology: that is, to replace highskill, slow, and costly artisanal work such as cloning and custom assays with computationally guided, automated, and standardized engineering procedures (Cameron et al., 2014; Chao et al., 2017). Although the industrialscale engineering potential of SynBio is far from fully realized at this point (Davies, 2019), it is very much here to stay and has begun to dictate major changes in biology education and training (National Research Council, 2015). Whether you are a trainee or a trainer, these trends are seriously worth considering for your future career's sake. A hallmark of SynBio is to rethink biological molecules, genes and proteins, as engineering "parts" that can be standardized, quantitatively characterized, and used to build a range of devices, much as standard resistors and capacitors are used as components in countless different electrical circuits (de Lorenzo and Schmidt, 2018). In the case of metabolic pathways, for instance, enzyme parts from plants or other organisms can be used as is and combined in novel ways to build

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Split-TALE: A TALE-Based Two-Component System for Synthetic Biology Applications in Planta

Cited by 16 publications

References 60 publications

Medicago TERPENE SYNTHASE 10 Is Involved in Defense Against an Oomycete Root Pathogen

Medicago TERPENE SYNTHASE 10 Is Involved in Defense Against an Oomycete Root Pathogen

Tomato MYB21 Acts in Ovules to Mediate Jasmonate-Regulated Fertility

Focus Issue Editorial: Synthetic Biology

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