A5014154862- CRISPR and Genetic Engineering
- Plant tissue culture and regeneration
- Transgenic Plants and Applications
- bioluminescence and chemiluminescence research
- Cell Image Analysis Techniques
- Animal Genetics and Reproduction
- Viral Infectious Diseases and Gene Expression in Insects
- Microbial Metabolic Engineering and Bioproduction
- Toxin Mechanisms and Immunotoxins
- Photoreceptor and optogenetics research
- Biosensors and Analytical Detection
- Botanical Research and Applications
- Plant nutrient uptake and metabolism
- Cellular Mechanics and Interactions
- Genetic Associations and Epidemiology
- Chemistry and Chemical Engineering
- Genomics and Rare Diseases
- Plant Virus Research Studies
- Modular Robots and Swarm Intelligence
- Electrophoretic Deposition in Materials Science
- Plant Stress Responses and Tolerance
- Engineering and Information Technology
- Insect Resistance and Genetics
- Legionella and Acanthamoeba research
- Photosynthetic Processes and Mechanisms
Baylor College of Medicine
2015-2023
Instituto de Biología Molecular y Celular de Plantas
2011-2017
Universitat Politècnica de València
2011-2016
Consejo Superior de Investigaciones Científicas
2013-2016
Plant synthetic biology aims to apply engineering principles plant genetic design. One strategic requirement of is the adoption common standardized technologies that facilitate construction increasingly complex multigene structures at DNA level while enabling exchange building blocks among bioengineers. Here, we describe GoldenBraid 2.0 (GB2.0), a comprehensive technological framework foster standard parts for biology. GB2.0 relies on use type IIS restriction enzymes assembly and proposes...
Synthetic Biology requires efficient and versatile DNA assembly systems to facilitate the building of new genetic modules/pathways from basic parts in a standardized way. Here we present GoldenBraid (GB), system based on type IIS restriction enzymes that allows indefinite growth reusable gene modules made pieces. The GB consists set four destination plasmids (pDGBs) designed incorporate multipartite assemblies standard combine them binarily build increasingly complex multigene constructs....
Abscisic acid (ABA) plays a crucial role in the plant’s response to both biotic and abiotic stress. Sustainable production of food faces several key challenges, particularly generation new varieties with improved water use efficiency drought tolerance. Different studies have shown potential applications Arabidopsis PYR/PYL/RCAR ABA receptors enhance plant resistance. Consequently functional characterization orthologous genes crops holds promise for agriculture. The full set tomato (Solanum...
Summary Betalains are tyrosine‐derived red‐violet and yellow pigments, found in plants only of the Caryophyllales order. Although much progress has been made recent years understanding betalain biosynthetic process, many questions remain open with regards to several proposed steps pathway. Most conspicuous by its absence is characterization first committed step pathway, namely 3‐hydroxylation tyrosine form l ‐3,4‐dihydroxyphenylalanine ( ‐ DOPA ). We used transcriptome analysis...
Modular DNA assembly simplifies multigene engineering in Plant Synthetic Biology. Furthermore, the recent adoption of a common syntax to facilitate exchange plant parts (phytobricks) is promising strategy speed up genetic engineering. Following this lead, here, we present platform for biodesign that incorporates functional descriptions phytobricks obtained under pre-defined experimental conditions, and systematically registers resulting information as metadata documentation. To handling...
Abstract Sensitive simultaneous assessment of multiple signaling pathways within the same cells requires orthogonal reporters that can assay over large dynamic ranges. Luciferases are such genetically encoded candidates due to their sensitivity, versatility, and cost-effectiveness. We expand luciferase multiplexing in post-lysis endpoint assays from two six. Light emissions distinguished by a combination distinct substrates emission spectra deconvolution. All six reporter units stitched...
In vitro derivation of pancreatic β-cells from human pluripotent stem cells holds promise as diabetes treatment. Despite recent progress, efforts to generate physiologically competent are still hindered by incomplete understanding the microenvironment's role in β-cell development and maturation. Here, we analyze mesenchymal endothelial primary weeks 9-20 fetal pancreas identify a time point-specific microenvironment that permits differentiation. Further, uncover unique factors guide...
Delivery of secretory immunoglobulin A (sIgA) to mucosal surfaces as a passive immunotherapy agent is promising strategy prevent infectious diseases. Recombinant sIgA production in plants requires the co-expression four transcriptional units encoding light chain (LC), heavy (HC), joining (JC) and component (SC). As way optimize plants, we tested combinatorial expression 16 versions human against VP8* rotavirus antigen Nicotiana benthamiana, using recently developed GoldenBraid multigene...
The power of Drosophila melanogaster as a model system relies on tractable germline genetic manipulations. Despite Drosophila's expansive genetics toolbox, such manipulations are still accomplished one change at time and depend predominantly phenotypic screening. We describe drug-based platform consisting four selection two counterselection markers, eliminating the need to screen for modified progeny. These markers work reliably individually or in combination produce specific outcomes....
Proanthocyanidins (PAs), or condensed tannins, are powerful antioxidants that remove harmful free oxygen radicals from cells. To engineer the anthocyanin and proanthocyanidin biosynthetic pathways to de novo produce PAs in two Nicotiana species, we incorporated four transgenes plant chassis. We opted perform a simultaneous transformation of genes linked multigenic construct rather than classical breeding retransformation approaches. generated GoldenBraid 2.0 containing Antirrhinum majus...
GoldenBraid is a rapid, modular, and robust cloning system used to assemble combine genetic elements. Dictyostelium amoebae represent an intriguing synthetic biological chassis with tractable applications in development, chemotaxis, bacteria-host interactions, allorecognition. We present as framework for Dictyostelium, including library of 250 DNA parts assemblies proof-of-concept strain that illustrates cAMP-chemotaxis four fluorescent reporters coded by one plasmid.
Abstract Many synthetic biologists have adopted methods based on Type IIS restriction enzymes and Golden Gate technology in their cloning procedures, as these enable the combinatorial assembly of modular elements a very efficient way following standard rules. GoldenBraid (GB) is Gate–based system that, addition, facilitates engineering large multigene constructs exchange DNA parts result its iterative scheme. GB was initially developed specifically for plant biology, it has been subsequently...
Abstract High‐throughput cell‐based screening assays are valuable tools in the discovery of chemical probes and therapeutic agents. Such designed to examine effects small compounds on targets, pathways, or phenotypes participating normal disease processes. While most measure single quantities, multiplexed seek address these limitations by obtaining multiple simultaneous measurements. The signals from such measurements should be independently detectable cover large dynamic ranges. Luciferases...
Abstract We recently described a drug‐based selectable and counterselectable genetic platform for the animal model system Drosophila melanogaster , consisting of four resistance two sensitivity markers that allow direct selection for, or counterselection against, desired genotype. This eliminates need to identify modified progeny by traditional laborious screening using dominant eye body color markers, white + yellow respectively. The permit animals G418 sulfate, puromycin HCl, blasticidin...
Multiplex experimentation that can assay multiple cellular signaling pathways in the same cells requires orthogonal genetically encoded reporters report over large dynamic ranges. Luciferases are cost-effective, versatile candidates whose output signals be sensitively detected a multiplex fashion. Commonly used dual luciferase reporter assays detect one is coupled to single pathway and second control for normalization purposes. We have expanded this approach hextuple on five control, each of...
Betalains are tyrosine-derived red-violet and yellow pigments found in plants only of the Caryophyllales order, which hold both scientific economic values. Their pH in-dependence high stability make them a natural pigment choice for food industries. strong antioxidant activities have prompted research into their potential health-promoting properties led to commercialization variety betalain-based dietary supplements. While biosynthetic process many colorants is well understood, questions...
Abstract Sensitive simultaneous assessment of multiple signaling pathways within the same cells requires orthogonal reporters that can assay over large dynamic ranges. Luciferases have arisen as possible genetically encoded candidates due to their sensitivity, versatility, and cost-effectiveness. Here, we expanded luciferase multiplexing in post-lysis endpoint assays from two towards six. Light emissions are distinguished by a combination distinct substrates emission spectra deconvolution....