A5051667493- Click Chemistry and Applications
- Advanced Fluorescence Microscopy Techniques
- Biotin and Related Studies
- Monoclonal and Polyclonal Antibodies Research
- Receptor Mechanisms and Signaling
- Advanced biosensing and bioanalysis techniques
- bioluminescence and chemiluminescence research
- Advanced Biosensing Techniques and Applications
- Photoreceptor and optogenetics research
- Chemical Synthesis and Analysis
- DNA Repair Mechanisms
- Tuberculosis Research and Epidemiology
- Photochromic and Fluorescence Chemistry
- Lipid Membrane Structure and Behavior
- Molecular Sensors and Ion Detection
- Luminescence and Fluorescent Materials
- DNA and Nucleic Acid Chemistry
- Cellular transport and secretion
- Photosynthetic Processes and Mechanisms
- Analytical Chemistry and Sensors
- Biosensors and Analytical Detection
- Enzyme Catalysis and Immobilization
- Pancreatic function and diabetes
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
École Polytechnique Fédérale de Lausanne
2016-2025
Max Planck Institute for Medical Research
2017-2025
University of Geneva
2011-2023
ORCID
2021
NCCR Chemical Biology - Visualisation and Control of Biological Processes Using Chemistry
2011-2019
Robert Bosch (Germany)
2019
Laboratoire de Chimie Moléculaire et Thioorganique
2019
Royal Australian Chemical Institute
2019
FORS – Swiss Centre of Expertise in the Social Sciences
2014-2018
Max Planck Society
2018
A general approach for the sequential labeling of fusion proteins O 6 -alkylguanine-DNA alkyltransferase (AGT) with different fluorophores in mammalian cells is presented. AGT localizations cell can be labeled specifically fluorophores, and fluorescence used applications such as multicolor analysis dynamic processes resonance energy transfer measurements. The facile access to a variety substrates well specificity reaction should make an important tool study protein function live cells.
Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used for imaging either toxic, require illumination with blue light or not compatible super-resolution microscopy, thereby limiting their utility. Here we describe a far-red stain, SiR-Hoechst, which displays minimal toxicity, is applicable different cell types and tissues, microscopy. The combination of these properties makes this probe powerful tool
Abstract The ability to specifically attach chemical probes individual proteins represents a powerful approach the study and manipulation of protein function in living cells. It provides simple, robust versatile imaging fusion wide range experimental settings. However, potential drawback detection using is fluorescence background from unreacted or nonspecifically bound probes. In this report we present design application novel fluorogenic for labeling SNAP‐tag an engineered variant human...
Here we present a far-red, silicon-rhodamine-based fluorophore (SiR700) for live-cell multicolor imaging. SiR700 has excitation and emission maxima at 690 715 nm, respectively. SiR700-based probes F-actin, microtubules, lysosomes, SNAP-tag are fluorogenic, cell-permeable, compatible with superresolution microscopy. In conjunction based on the previously introduced carboxy-SiR650, permit microscopy in thus significantly expanding our capacity imaging living cells.
A cell-based phenotypic screen for inhibitors of biofilm formation in mycobacteria identified the small molecule TCA1, which has bactericidal activity against both drug-susceptible and -resistant Mycobacterium tuberculosis (Mtb) sterilizes Mtb vitro combined with rifampicin or isoniazid. In addition, TCA1 nonreplicating is efficacious acute chronic infection mouse models alone Transcriptional analysis revealed that down-regulates genes known to be involved persistence. Genetic affinity-based...
Rhodamines are the most important class of fluorophores for applications in live-cell fluorescence microscopy. This is mainly because rhodamines exist a dynamic equilibrium between fluorescent zwitterion and nonfluorescent but cell-permeable spirocyclic form. Different imaging require different positions this equilibrium, an adjustment poses challenge design suitable probes. We describe here how conversion ortho-carboxy moiety given rhodamine into substituted acyl benzenesulfonamides...
Abstract Self-labeling protein tags such as HaloTag are powerful tools that can label fusion proteins with synthetic fluorophores for use in fluorescence microscopy. Here we introduce variants either increased or decreased brightness and lifetime compared HaloTag7 when labeled rhodamines. Combining these enabled live-cell multiplexing of three cellular targets one spectral channel using a single fluorophore the generation lifetime-based biosensor. Additionally, brightest variant showed up to...
The self-labeling protein tags (SLPs) HaloTag7, SNAP-tag, and CLIP-tag allow the covalent labeling of fusion proteins with synthetic molecules for applications in bioimaging biotechnology. To guide selection an SLP–substrate pair provide guidelines design substrates, we report a systematic comparative study kinetics substrate specificities CLIP-tag. HaloTag7 reaches almost diffusion-limited rate constants certain rhodamine which are more than 2 orders magnitude higher those SNAP-tag...
Abstract The quality and application of super-resolution fluorescence imaging greatly lie in the dyes’ properties, including photostability, brightness, Stokes shift. Here we report a synergistic strategy to simultaneously improve such properties regular fluorophores. Introduction quinoxaline motif with fine-tuned electron density conventional rhodamines generates new dyes vibration structure inhibited twisted-intramolecular-charge-transfer (TICT) formation synchronously, thus increasing...
The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, irreversible attachment probe to precludes imaging applications that require transient binding comes risk photobleaching. Here, we introduce exchangeable ligands fluorescence (xHTLs) reversibly bind can be coupled rhodamines different colors. In stimulated emission depletion (STED) microscopy, exchange xHTLs allows reduced photobleaching as...
Fluorescent biosensors enable the study of cell physiology with spatiotemporal resolution; yet, most suffer from relatively low dynamic ranges. Here, we introduce a family designed Förster resonance energy transfer (FRET) pairs near-quantitative FRET efficiencies based on reversible interaction fluorescent proteins fluorescently labeled HaloTag. These enabled straightforward design for calcium, ATP and NAD+ unprecedented The color each these can be readily tuned by changing either protein or...
Recordings of the physiological history cells provide insights into biological processes, yet obtaining such recordings is a challenge. To address this, we introduce method to record transient cellular events for later analysis. We designed proteins that become labeled in presence both specific activity and fluorescent substrate. The recording period set by substrate, whereas controls degree labeling. use distinguishable substrates enabled successive periods activity. recorded...
The specific and covalent labeling of fusion proteins with synthetic molecules opens up new ways to study protein function in the living cell. Here we present a novel method that allows for exclusive extracellular on surfaces live cells large variety including fluorophores, ligands, or quantum dots. approach is based acyl carrier through post-translational modification catalyzed by phosphopantetheine transferase. specificity versatility should allow it become an important tool studying...