A5021853457- Pluripotent Stem Cells Research
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- 3D Printing in Biomedical Research
- Virus-based gene therapy research
- Advanced biosensing and bioanalysis techniques
- Single-cell and spatial transcriptomics
- Neuroinflammation and Neurodegeneration Mechanisms
- Viral Infectious Diseases and Gene Expression in Insects
- Phagocytosis and Immune Regulation
- Animal Genetics and Reproduction
- Receptor Mechanisms and Signaling
- Prenatal Screening and Diagnostics
- Barrier Structure and Function Studies
- Cell Image Analysis Techniques
- Renal and related cancers
- Alzheimer's disease research and treatments
- Cholinesterase and Neurodegenerative Diseases
- Gene Regulatory Network Analysis
- Immune cells in cancer
- Lysosomal Storage Disorders Research
- Calcium signaling and nucleotide metabolism
- Inflammation biomarkers and pathways
- Tuberous Sclerosis Complex Research
- Hedgehog Signaling Pathway Studies
Roche (Switzerland)
2013-2024
Merck (Germany)
2024
Life & Brain (Germany)
2006-2010
Hertie Foundation
2006-2010
University of Bonn
2006-2010
Cell-penetrating peptides (CPPs) are capable of introducing a wide range cargoes into living cells. Descriptions the internalization process vary from energy-independent cell penetration membranes to endocytic uptake. To elucidate whether mechanism entry CPP constructs might be influenced by properties cargo, we used time lapse confocal microscopy analysis mammalian cells directly compare uptake well-studied TAT fused protein (>50 amino acids) or peptide (<50 cargo. We also analyzed various...
Tyrosine site-specific recombinases (SSRs) including Cre and FLP are essential tools for DNA genome engineering. has long been recognized as the best SSR engineering, particularly in mice. Obtaining another that is good will be a valuable addition to genomic toolbox. To this end, we have developed validated reagents Dre-rox system. These include an Escherichia coli-inducible expression vector based on temperature-sensitive pSC101 plasmid, mammalian CAGGs promoter, rox-lacZ reporter embryonic...
Background In preimplantation mammalian development the transcription factor Sox2 (SRY-related HMG-box gene 2) forms a complex with Oct4 and functions in maintenance of self-renewal pluripotent inner cell mass (ICM). Previously it was shown that Sox2−/− embryos die soon after implantation. However, maternal transcripts may mask an earlier phenotype. We investigated whether is involved controlling fate decisions at stage. Methods Findings addressed question role for using RNAi, which removes...
Reproducibility in molecular and cellular studies is fundamental to scientific discovery. To establish the reproducibility of a well-defined long-term neuronal differentiation protocol, we repeated comparison same two iPSC lines across five distinct laboratories. Despite uncovering acceptable variability within individual laboratories, detect poor cross-site differential gene expression signature between these lines. Factor analysis identifies laboratory as largest source variation along...
The blood brain barrier (BBB) has evolved unique characteristics such as dense coverage of the endothelial cells by pericytes and interactions with astrocytes through perivascular endfeet. We study BBB formation in a 3-dimensional multicellular spheroid system human primary (hpBECs), (hpPs) (hpAs). show for first time that hpBECs, hpPs hpAs spontaneously self-organize into defined structure which recapitulates complex arrangement individual cell types structure. Pericytes play crucial role...
Highlights•TSC2 deletion causes gene-dosage-dependent mTORC1 hyperactivity in neurodevelopment•TSC2 alterations pathways and genes associated with autism•TSC2+/− TSC2−/− human neurons show stage-specific cellular synaptic defects•mTORC1 inhibition corrects defects independently of early neurodevelopmentSummaryHyperfunction the pathway has been idiopathic syndromic forms autism spectrum disorder (ASD), including tuberous sclerosis, caused by loss either TSC1 or TSC2. It remains largely...
Tissue-resident macrophages are key players in inflammatory processes, and their activation functionality crucial health disease. Numerous diseases associated with alterations homeostasis or dysregulation of the innate immune system, including allergic reactions, autoimmune diseases, cancer. Macrophages a prime target for drug discovery due to major regulatory role Currently, main sources used therapeutic compound screening primary cells isolated from blood tissue immortalized neoplastic...
Generating human podocytes in vitro could offer a unique opportunity to study diseases. Here, we describe simple and efficient protocol for obtaining functional from induced pluripotent stem cells. Cells were exposed three-step protocol, which their differentiation into intermediate mesoderm, then nephron progenitors and, finally, mature podocytes. After differentiation, cells expressed the main podocyte markers, such as synaptopodin, WT1, α-Actinin-4, P-cadherin nephrin at protein mRNA...
The blood-retina barrier and blood-brain (BRB/BBB) are selective semipermeable critical for supporting protecting central nervous system (CNS)-resident cells. Endothelial cells (ECs) within the BRB/BBB tightly coupled, express high levels of Claudin-5 (CLDN5), a junctional protein that stabilizes ECs, important proper neuronal function. To identify novel CLDN5 regulators (and ultimately EC stabilizers), we generated CLDN5-P2A-GFP stable cell line from human pluripotent stem (hPSCs), directed...
Angelman syndrome (AS) is a neurodevelopmental disorder caused by the loss of maternal UBE3A, ubiquitin protein ligase E3A. Here, we study neurons derived from patients with AS and neurotypical individuals, reciprocally modulate UBE3A using antisense oligonucleotides. Unbiased proteomics reveal proteins that are regulated in disease-specific manner, including PEG10, retrotransposon-derived GAG protein. PEG10 increase, but not RNA, dependent on proteasome function. binds to both RNA...
Microglia are key in the homeostatic well-being of brain and microglial dysfunction has been implicated neurodegenerative disorders such as Alzheimer's disease (AD). Due to many limitations study microglia situ or isolated for large scale drug discovery applications, there is a high need develop robust scalable human cellular models with reliable translatability disease. Here we describe generation microglia-like cells from induced pluripotent stem (iPSC) distinct phenotypes mechanistic...
Abstract Endothelial cells (ECs) display remarkable plasticity during development before becoming quiescent and functionally mature. EC maturation is directed by several known transcription factors (TFs), but the specific set of TFs responsible for promoting high-resistance barriers, such as blood-brain barrier (BBB), have not yet been fully defined. Using expression mRNA data from published studies on ex vivo ECs central nervous system (CNS), we predicted that induce properties in BBB. We...
Abstract Removal of apoptotic cells by phagocytes (also called efferocytosis) is a crucial process for tissue homeostasis. Professional express plethora surface receptors enabling them to sense and engulf cells, thus avoiding persistence dead cellular debris their consequent effects. Dysregulation efferocytosis thought lead secondary necrosis associated inflammation immune activation. Efferocytosis in primarily murine macrophages dendritic has been shown require TAM RTKs, with MERTK AXL...
Mucopolysaccharidoses are inherited metabolic disorders caused by the deficiency in lysosomal enzymes required to break down glycosaminoglycans. Accumulation of glycosaminoglycans leads progressive, systemic degenerative disease. The central nervous system is particularly affected, resulting developmental delays, neurological regression, and early mortality. Current treatments fail adequately address defects. Here we explore potential human induced pluripotent stem cell (hiPSC)-derived...
Therapeutic approaches to fight Alzheimer's disease include anti-Amyloidβ (Aβ) antibodies and secretase inhibitors. However, the blood-brain barrier (BBB) limits brain exposure of biologics chemical space for small molecules be BBB permeable. The Brain Shuttle (BS) technology is capable shuttling large into brain. This allows new types therapeutic modalities engineered optimal efficacy on molecular target in independent penetrating properties. To this end, we designed BACE1 peptide...
Abstract Continuous expression of Cre recombinase has the potential to yield toxic side effects in various cell types, thereby limiting applications Cre/loxP system for conditional mutagenesis. In this study, we investigate protein transduction overcome limitation. COS‐7, CV1‐5B, and mouse embryonic stem (ES) cells treated with cell‐permeant (HTNCre) maintain a normal growth behavior employing concentrations sufficient induce recombination more than 90% cells, whereas continuous application...
Purpose Human pluripotent stem cell (hPSC)‐derived cellular models have great potential to enable drug discovery and improve translation of preclinical insights the clinic. We developed a hPSC‐derived neural precursor model for studying early events in human brain development. present protein‐level characterization this model, using multiplexed SRM approach, establish reproducibility physiological relevance; essential prerequisites utilization neuronal development phenotypic screening‐based...
Abstract Combined application of DNA recombinases Cre and FLP enables tightly controlled independent and/or sequential gene regulations. However, in practice, such dual recombinase strategies are hampered by the comparably low efficiency recombinase. Here, we present engineering a recombinant cell-permeant protein (TAT-FLP) that induces recombination &gt;75% fibroblasts mouse as well human embryonic stem (ES) cells. We show TAT-FLP ideally complements strength for genetic exemplified...
Abstract Due to their broad differentiation potential, pluripotent stem cells (PSCs) offer a promising approach for generating relevant cellular models various applications. While human PSC-based are already advanced, similar systems non-human primates (NHPs) still lacking. However, as NHPs the most appropriate animals evaluating safety of many novel pharmaceuticals, availability in vitro would be extremely useful bridge gap between and animal models. Here, we present NHP endothelial cell...
The protein transduction technique enables the direct delivery of biologically active material into mammalian cells [for review see 1,2]. For this one can make use translocating ability so-called cell penetrating peptides (CPPs), also designated as domains (PTDs). TAT-CPP derived from human immunodeficiency virus type 1 (HIV-1) Tat (trans-activator transcription) has been widely used. positively charged TAT promotes permeability thereby overcoming barriers cellular membrane by endocytosis...