A5014052655- Genomics and Chromatin Dynamics
- Molecular Junctions and Nanostructures
- CRISPR and Genetic Engineering
- Protein Degradation and Inhibitors
- Alzheimer's disease research and treatments
- Advanced biosensing and bioanalysis techniques
- DNA and Nucleic Acid Chemistry
- RNA Research and Splicing
- Biochemical and Structural Characterization
- Cellular transport and secretion
- Ubiquitin and proteasome pathways
- Virus-based gene therapy research
- Epigenetics and DNA Methylation
- Amino Acid Enzymes and Metabolism
- Nanopore and Nanochannel Transport Studies
- Botulinum Toxin and Related Neurological Disorders
- RNA and protein synthesis mechanisms
- Nutrition, Genetics, and Disease
- RNA modifications and cancer
- Advanced Fluorescence Microscopy Techniques
- CAR-T cell therapy research
- Fungal and yeast genetics research
- Viral Infectious Diseases and Gene Expression in Insects
- Animal Genetics and Reproduction
- Protein Structure and Dynamics
University of Fribourg
2022-2024
Adolphe Merkle Institute
2022-2024
École Polytechnique Fédérale de Lausanne
2014-2022
Abstract The dynamic architecture of chromatin fibers, a key determinant genome regulation, is poorly understood. Here, we employ multimodal single-molecule Förster resonance energy transfer studies to reveal structural states and their interconversion kinetics in fibers. We show that nucleosomes engage short-lived (micro- milliseconds) stacking interactions with one neighbors. This results discrete tetranucleosome units distinct interaction registers interconvert within hundreds...
Abstract Multivalent interactions between effector proteins and histone post-translational modifications are an elementary mechanism of dynamic chromatin signalling. Here we elucidate the how heterochromatin protein 1α (HP1α), a multivalent effector, is efficiently recruited to silent state (marked by trimethylated H3 at Lys9, H3K9me3) while remaining highly dynamic. Employing chemically defined nucleosome arrays together with single-molecule total internal reflection fluorescence microscopy...
Chromatin recruitment of effector proteins involved in gene regulation depends on multivalent interaction with histone post-translational modifications (PTMs) and structural features the chromatin fiber. Due to complex interactions involved, it is currently not understood how effectors dynamically sample landscape. Here, we dissect dynamic a family effectors, heterochromatin protein 1 (HP1) proteins, using single-molecule fluorescence imaging computational modeling. We show that three human...
Early-stage aggregates of amyloid-forming proteins, specifically soluble oligomers, are implicated in neurodegenerative diseases such as Alzheimer's disease, Parkinson's and Huntington's disease. Protein aggregation is typically monitored by fluorescence using the amyloid-binding fluorophore thioflavin T (ThT). Thioflavin interacts, however, preferentially with fibrillar amyloid structures rather than soluble, early-stage aggregates. In contrast, two fluorophores, aminonaphthalene...
Aerosol-mediated delivery of nano-based therapeutics to the lung has emerged as a promising alternative for treatment and prevention diseases. Superparamagnetic iron oxide nanoparticles (SPIONs) have attracted significant attention such applications due their biocompatibility magnetic properties. However, information is lacking about characteristics nebulized SPIONs use therapeutic aerosol. To address this need, we conducted physicochemical characterization Rienso, SPION-based formulation...
KAP1 (KRAB domain–associated protein 1) plays a fundamental role in regulating gene expression mammalian cells by recruiting different transcription factors and altering the chromatin state. In doing so, acts both as platform for macromolecular interactions an E3 small ubiquitin modifier ligase. This work sheds light on overall organization of full-length combining solution scattering data, integrative modeling, single-molecule experiments. We show that is elongated antiparallel dimer with...
Single-molecule measurements provide detailed mechanistic insights into molecular processes, for example in genome regulation where DNA access is controlled by nucleosomes and the chromatin machinery. However, real-time single-molecule observations of nuclear factors acting on defined substrates are challenging to perform quantitatively reproducibly. Here we present XSCAN (multiple
Abstract KAP1 (KRAB-domain associated protein 1) plays a fundamental role in regulating gene expression mammalian cells by recruiting different transcription factors and altering the chromatin state. In doing so, acts both as platform for macromolecular interactions an E3 SUMO ligase. This work sheds light on overall organization of full-length combining solution scattering diffraction data, integrative modeling single-molecule experiments. We show that is elongated antiparallel dimer with...
Formation of Tau protein aggregates in neurons is a pathological hallmark several neurodegenerative diseases, including Alzheimer's disease. Fluorescently labeled therefore useful to study the aggregation these proteins and identify potential therapeutic targets. Conventionally, cysteine residues are used for labeling proteins; however, full-length isoform contains two microtubule-binding region, which implicated by forming intermolecular disulfide bonds. To prevent fluorescent label from...
Viral vectors for gene therapy, such as recombinant adeno-associated viruses, are produced in human embryonic kidney (HEK) 293 cells. However, the presence of SV40 T-antigen-encoding CDS SV40GP6 and SV40GP7 HEK293T genome raises safety issues when these cells used manufacturing clinical purposes. We developed a new T-antigen-negative HEK cell line from ExcellGene's proprietary HEKExpress,® using CRISPR-Cas9 strategy. obtained high number clonally-derived populations all them were...
Many proteins undergo conformational changes during their activity. A full understanding of the function these can only be obtained if different conformations and transitions between them monitored in aqueous solution, with adequate temporal resolution and, ideally, on a single-molecule level. Interrogating dynamics single remains, however, exquisitely challenging typically requires site-directed chemical modification combined rigorous minimization possible artifacts. These obstacles limit...
Keywords: Summer school ; Epigenetics Chemical biology Cell Chromatin Structure Reference EPFL-ARTICLE-213778doi:10.2533/chimia.2015.624View record in Web of Science Record created on 2015-11-26, modified 2017-05-12
Viral vectors for gene therapy, such as recombinant Adeno-Associated Viruses (rAAV), are produced in Human Embryonic Kidney (HEK) 293 cells. However, the presence of SV40 T-antigen-encoding CDS SV40GP6 and SV40GP7 HEK293T genome raises safety issues when these cells used manufacturing clinical purposes. We developed a new T-antigen-negative HEK cell line from ExcellGene’s proprietary HEKExpress®, using CRISPR-Cas9 strategy. obtained high number clonally-derived populations all them were...
Fluorescence microscopy images of amyloid oligomers, protofibrils and fibrils formed by native labelled Ta u protein.Left: Native protein labeled with thioflavin T (ThT) in solution.Right: Alexa647-labeled without ThT solution.