Core promoter sequence in yeast is a major determinant of expression level. GAL4 GFP enhancer trap lines for analysis of stomatal guard cell development and gene expression. GAL4-GFP enhancer trap lines for genetic manipulation of lateral root development in Arabidopsis thaliana. GFP variants for multispectral imaging of living cells. A synthetic biology framework for programming eukaryotic transcription functions. Designable DNA-binding domains enable construction of logic circuits in mammalian cells. Benchmarking of TALE- and CRISPR/dCas9-based transcriptional regulators in mammalian cells for the construction of synthetic genetic circuits. Tunable and multifunctional eukaryotic transcription factors based on CRISPR/Cas. Complex signal processing in synthetic gene circuits using cooperative regulatory assemblies. A library of synthetic transcription activator-like effector-activated promoters for coordinated orthogonal gene expression in plants. Synchronous long-term oscillations in a synthetic gene circuit. Production of benzylisoquinoline alkaloids in Saccharomyces cerevisiae. Biotechnology and synthetic biology approaches for metabolic engineering of bioenergy crops. Our findings demonstrate that these tools provide variation in transcriptional output while enabling the concerted expression of multiple genes in a tissue-specific and environmentally responsive manner, providing a basis for generating complex genetic circuits that process endogenous and environmental stimuli. This characterization of contributing genetic elements that dictate gene expression represents a foundation for the rational design of refined synthetic regulators. By leveraging orthogonal regulatory systems from Saccharomyces spp., we have developed a strategy for the design of synthetic activators, synthetic repressors, and synthetic promoters and have validated their use in Nicotiana benthamiana and Arabidopsis thaliana. We have successfully engineered a library of synthetic transcriptional regulators that modulate expression strength in planta. However, most efforts are hindered by a lack of characterized tools that allow for reliable and targeted expression of transgenes. Agricultural biotechnology strategies often require the precise regulation of multiple genes to effectively modify complex plant traits.
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