A5037627898- Click Chemistry and Applications
- Advanced Fluorescence Microscopy Techniques
- Advanced Biosensing Techniques and Applications
- Monoclonal and Polyclonal Antibodies Research
- Cell Image Analysis Techniques
- Advanced Electron Microscopy Techniques and Applications
- Neurogenesis and neuroplasticity mechanisms
- Advanced biosensing and bioanalysis techniques
- Chemical Synthesis and Analysis
- Biotin and Related Studies
- Neuroinflammation and Neurodegeneration Mechanisms
- RNA and protein synthesis mechanisms
- RNA regulation and disease
- Parkinson's Disease Mechanisms and Treatments
- Multiple Sclerosis Research Studies
- Cytomegalovirus and herpesvirus research
- HIV Research and Treatment
- Signaling Pathways in Disease
- RNA Interference and Gene Delivery
- Molecular Biology Techniques and Applications
- Nerve injury and regeneration
- Optical Coherence Tomography Applications
- Digital Holography and Microscopy
- Skin and Cellular Biology Research
- Zebrafish Biomedical Research Applications
University of Tübingen
2016-2025
Senckenberg Centre for Human Evolution and Palaeoenvironment
2016-2019
European Molecular Biology Laboratory
2014-2017
Ludwig-Maximilians-Universität München
2007-2016
European Molecular Biology Organization
2015-2016
European Bioinformatics Institute
2013-2015
European Molecular Biology Laboratory
2015
Abstract The growing demands of advanced fluorescence and super‐resolution microscopy benefit from the development small highly photostable fluorescent probes. Techniques developed to expand genetic code permit residue‐specific encoding unnatural amino acids (UAAs) armed with novel clickable chemical handles into proteins in living cells. Here we present design new UAAs bearing strained alkene side chains that have improved biocompatibility stability for attachment tetrazine‐functionalized...
Abstract Modern light microscopy, including super-resolution techniques, has brought about a demand for small labeling tags that bring the fluorophore closer to target. This challenge can be addressed by unnatural amino acids (UAAs) with bioorthogonal click chemistry. The minimal size of UAA and possibility couple fluorophores directly protein interest single-residue precision in living cells make unique. Here, we establish primary neurons use it fixed-cell, live-cell, dual-color...
Super-resolution microscopy (SRM) greatly benefits from the ability to install small photostable fluorescent labels into proteins. Genetic code expansion (GCE) technology addresses this demand, allowing introduction of labeling sites, in form uniquely reactive noncanonical amino acids (ncAAs), at any residue a target protein. However, low incorporation efficiency ncAAs and high background fluorescence limit its current SRM applications. Redirecting subcellular localization pyrrolysine-based...
Abstract Parkinson’s disease-associated kinase LRRK2 has been linked to IFN type II (IFN-γ) response in infections and dopaminergic neuronal loss. However, whether how synergizes with IFN-γ remains unclear. In this study, we employed neurons microglia differentiated from patient-derived induced pluripotent stem cells carrying G2019S, the most common mutation. We show that enhances G2019S-dependent negative regulation of AKT phosphorylation NFAT activation, thereby increasing vulnerability...
Abstract trans ‐Cyclooctene groups incorporated into proteins via non‐canonical amino acids (ncAAs) are emerging as specific handles for bioorthogonal chemistry. Here, we present a highly improved synthetic access to the axially and equatorially linked ‐cyclooct‐2‐ene isomers ( 1 , b ). We further show that connected isomer has half‐life about 10 times higher than equatorial reacts with tetrazines much faster, determined by stopped‐flow experiments. The properties resulted in different...
Abstract Oligodendrocyte damage is a central event in the pathogenesis of common neuroinflammatory condition, multiple sclerosis (MS). Where and how oligodendrocyte initiated MS not completely understood. Here, we use combination light electron microscopy techniques to provide dynamic highly resolved view lesions. We show that both its animal model structural at myelin sheaths only later spreads cell body. Early itself characterized by formation local out-foldings—‘myelinosomes’—, which are...
Introduction of bioorthogonal functionalities (e.g., trans-cyclooctene-TCO) into a protein interest by site-specific genetic encoding non-canonical amino acids (ncAAs) creates uniquely targetable platforms for fluorescent labeling schemes in combination with tetrazine-functionalized dyes. However, an intracellular is usually compromised high background, arising from the hydrophobicity ncAAs; this typically compensated hours-long washout to remove excess ncAAs cellular interior. To overcome...
Abstract Die steigende Nachfrage nach fortschrittlicher Fluoreszenzmikroskopie und hochauflösenden bildgebenden Verfahren (SRM) erfordert die Entwicklung neuer Methoden zum Markieren von Proteinen mit kleinen, photostabilen Fluorophoren, wenn möglich mehrfarbig. Technologie zur Erweiterung des genetischen Codes ermöglicht den positionsspezifischen Einbau einer nichtnatürlichen Aminosäure (UAS) chemisch biokompatibel modifizierbaren funktionellen Gruppen in lebenden Zellen. Wir präsentieren...
Abstract Site‐specific labeling of biomolecules is rapidly advancing due to the discovery novel mutually orthogonal reactions. Quantum chemistry studies have also increased our understanding their relative rates, although these until now been based on highly simplified reactants. Here we examine a set strain‐promoted click‐type cycloaddition reactions n ‐propyl azide, 3‐benzyl tetrazine and 3‐benzyl‐6‐methyl with cyclooctenes/ynes, in which aim address all relevant structural details Our...
Live cell super-resolution microscopy (SRM) imaging is often limited by the lack of optimal fluorescent labelling approaches. Here, we report two new chemogenetic approaches allowing far-red fluorescence detection to label intracellular protein targets achieve background free SRM images in fixed and live mammalian cells. These methods are based on Fluorescence-activating Absorption Shifting Tag (FAST), a small monomeric tag that forms reversible assembly with various rhodanine fluorophores....
Synthetic procedures for the construction of fluorogenic azido-labels were developed. Photophysical properties elaborated by experimental and theoretical investigations. Of newly synthesized bioorthogonally applicable dyes two selected on basis their performance further subjected to in vitro vivo studies. Both tags exhibited excellent as aqueous medium, azide form is virtually non-fluorescent, while "clicked" triazole congeners showed intense fluorescence. One these labels a very large...
Synthesis of a set new, azide bearing, biorthogonally applicable fluorogenic dyes with large Stokes shifts is presented herein. To assess the performance these new we have labeled genetically modulated, cyclooctyne-bearing protein in lysate medium. Studies showed that labels produce specific signal minimal background fluorescence. We also provide theoretical insights into design such labels.
The advent of super-resolution microscopy (nanoscopy) has set high standards for fluorescence tagging. Fluorescent proteins (FPs) are convenient tags in conventional imaging, but their use nanoscopy been questioned due to relatively large size and propensity form multimers. Here, we compared the nanoscale organization with or without FP by introducing unnatural amino acid propargyl-L-lysine (PRK) 26 known multimolecular arrangements into FP-tagged variants. We revealed coupling synthetic...
Small synthetic fluorophores are in many ways superior to fluorescent proteins as labels for imaging. A major challenge is use them a protein-specific labeling living cells. Here, we report on our of noncanonical amino acids that genetically encoded via the pyrrolysyl-tRNA/pyrrolysyl-RNA synthetase pair at artificially introduced TAG codons recoded E. coli strain. The strain lacking endogenous and TAG-specific release factor RF1. contain bioorthogonal groups can be clicked externally...