A5064965593- Virus-based gene therapy research
- Herpesvirus Infections and Treatments
- RNA Interference and Gene Delivery
- Viral Infections and Immunology Research
- Viral Infectious Diseases and Gene Expression in Insects
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- Neuropeptides and Animal Physiology
- Animal Virus Infections Studies
- Ubiquitin and proteasome pathways
- Receptor Mechanisms and Signaling
- Neuroendocrine regulation and behavior
- RNA modifications and cancer
- Viral gastroenteritis research and epidemiology
- Cancer-related Molecular Pathways
- RNA Research and Splicing
- Parvovirus B19 Infection Studies
- Adipose Tissue and Metabolism
- Cytomegalovirus and herpesvirus research
- Bacteriophages and microbial interactions
- Cancer therapeutics and mechanisms
- Cancer Research and Treatments
- Cancer-related molecular mechanisms research
- Retinoids in leukemia and cellular processes
- Cancer, Stress, Anesthesia, and Immune Response
Charité - Universitätsmedizin Berlin
2010-2022
Freie Universität Berlin
1999-2022
Humboldt-Universität zu Berlin
2018-2022
MSB Medical School Berlin
2011-2016
Medical University of Graz
2009
California Institute for Medical Research
2002
German Cancer Research Center
1993-1999
DKFZ-ZMBH Alliance
1993-1999
Max Planck Institute of Biochemistry
1999
TU Wien
1993
Background— RNA interference (RNAi) has the potential to be a novel therapeutic strategy in diverse areas of medicine. Here, we report on targeted RNAi for treatment heart failure, an important disorder humans that results from multiple causes. Successful failure is demonstrated rat model transaortic banding by targeting phospholamban, key regulator cardiac Ca 2+ homeostasis. Whereas gene therapy rests recombinant protein expression as its basic principle, uses regulatory RNAs achieve...
Using immunofluorescence and in situ hybridization techniques, we studied the intracellular localization of adeno-associated virus type 2 (AAV-2) Rep proteins, VP DNA during course an AAV-2/adenovirus coinfection. In early stage, proteins showed a punctate distribution pattern over nuclei infected cells, reminiscent replication foci. At this no capsid were detectable. later stages, distributed more homogeneously nuclear interior finally became redistributed into clusters slightly enriched at...
Human Topors, which was originally identified as cellular binding partner of DNA topoisomerase I and p53, has recently been shown to function an ubiquitin E3 ligase for p53 in a manner dependent on its N′‐terminally located RING finger. Here, we demonstrate that Topors also enhances the conjugation small ubiquitin‐like modifier 1 (SUMO‐1) vivo reconstituted vitro system. The SUMO‐1 activity does not depend upon finger motif. In HeLa cells, induced sumoylation accompanied by increase...
Anxiety is integrated in the amygdaloid nuclei and involves interplay of amygdala various other areas brain. Neuropeptides play a critical role regulating this process. Neuropeptide Y (NPY), 36 aa peptide, highly expressed amygdala. It exerts potent anxiolytic effects through cognate postsynaptic Y1 receptors, but augments anxiety presynaptic Y2 receptors. To identify precise anatomical site(s) Y2-mediated anxiogenic action, we investigated effect site-specific deletion gene on...
Scalable and genetically stable recombinant adeno-associated virus (rAAV) production systems combined with facile adaptability for an extended repertoire of AAV serotypes are required to keep pace the rapidly increasing clinical demand. For scalable high-titer full range rAAV 1-12, we developed OneBac, consisting insect Sf9 cell lines harboring silent copies AAV1-12 rep cap genes induced upon infection a single baculovirus that also carries genome. burst sizes reach up 5 × 10(5)...
The proteins encoded by the adeno-associated virus type 2 (AAV-2) rep and cap genes obtained during a productive infection of HeLa cells with AAV-2 adenovirus were fractionated according to solubility, cellular localization, sedimentation properties. majority Rep Cap accumulated in nucleus, where they distributed into soluble an insoluble fraction. Analysis nuclear fraction capsid sucrose density gradients showed that formed at least three steady-state pools: monomer pool sedimenting about...
The four Rep proteins encoded by adeno-associated virus type 2 (AAV-2) inhibit transcription of their own promoters and several heterologous promoters. To gain insight into the molecular mechanism Rep-mediated repression, we studied effects on accumulation mRNA transcribed from human papillomavirus 18 upstream regulatory region HPV18 URR, immunodeficiency long terminal repeat, AAV-2 p5 p19 transient transfection experiments in HeLa cells. We observed a distinct contribution C- N-terminal...
Despite its strong host tropism for erythroid progenitor cells, human parvovirus B19 (B19V) can also infect a variety of additional cell types. Acute and chronic inflammatory cardiomyopathies have been associated with high prevalence B19V DNA in endothelial cells the myocardium. To elucidate mechanisms uptake into endothelium, we first analyzed surface expression well-characterized primary receptor P antigen putative coreceptors α5β1 integrins Ku80 on permanent cells. The pattern attachment...
Temporal lobe epilepsy (TLE) is the most frequent form of focal epilepsies and generally associated with malfunctioning hippocampal formation. Recently, a preferential loss parvalbumin (PV) neurons has been observed in subiculum TLE patients animal models TLE. To demonstrate possible causative role defunct PV generation TLE, we permanently inhibited GABA release selectively from ventral by injecting viral vector expressing tetanus toxin light chain male mice. Subsequently, mice were...
ABSTRACT The nonstructural adeno-associated virus type 2 Rep proteins are known to control viral replication and thus provide the single-stranded DNA genomes required for packaging into preformed capsids. In addition, complexes between capsids have previously been observed in course of productive infections. Such interpreted as genome-linked molecules associated with capsid upon successful encapsidation. Here we demonstrate via coimmunoprecipitation, cosedimentation, yeast two-hybrid...
Adeno-associated virus type 2 (AAV) is known to establish latency by preferential integration in human chromosome 19q13.42. The AAV non-structural protein Rep appears target a site called AAVS1 simultaneously binding Rep-binding sites (RBS) present on the genome and within AAVS1. In absence of Rep, as case with vectors, chromosomal rare random. For genome-wide survey wildtype linker-selection-mediated (LSM)-PCR strategy was designed retrieve AAV-chromosomal junctions. DNA sequence...
Scalable production of recombinant adeno-associated virus vectors (rAAV) in baculovirus-infected Sf9 cells yields high burst sizes but variable infectivity rates per packaged AAV vector genome depending on the chosen serotype. Infectivity are particularly low for rAAV5 vectors, based genetically most divergent In this study we describe key improvements OneBac system generation whose manufacturing has been unsatisfactory all current insect cell-based systems. The expression strategy AAV5...
ABSTRACT Seroepidemiology shows that infections with adeno-associated virus (AAV) are widespread, but diverse AAV serotypes isolated from humans or nonhuman primates have so far not been proven to be causes of human disease. In view the increasing success AAV-derived vectors in gene therapy, definition vivo sites wild-type persistence and clinical consequences its reactivation is becoming increasingly urgent. Here, we identify presumed cell type for host by highly sensitive PCRs developed...
ABSTRACT Adeno-associated virus (AAV) is recognized for its bipartite life cycle with productive replication dependent on coinfection adenovirus (Ad) and AAV latency being established in the absence of a helper virus. The shift from latent to Ad-dependent mostly regulated at transcriptional level. current transcription map displays highly expressed transcripts as found upon Ad. So far, have only been characterized plus strand single-stranded DNA genome. minus assumed not be transcribed....
ABSTRACT The adeno-associated virus type 2 (AAV-2) Rep78/Rep68 regulatory proteins are pleiotropic effectors of viral and cellular DNA replication, transformation by oncogenes, homologous heterologous gene expression. To search for involved in mediating these functions, we used Rep68 as bait the yeast two-hybrid system identified transcriptional coactivator PC4 a Rep interaction partner. has been shown to mediate activation variety sequence-specific transcription factors vitro. amino acids...
Adeno-associated virus type 2 (AAV-2) gene expression is tightly controlled by functions of the helper as well products its own viral rep gene. Double-immunofluorescence studies Rep and VP protein in cells coinfected with AAV-2 adenovirus showed that a large proportion these expressed Rep78 Rep52 but no capsid proteins. The percentage Rep78/Rep52- protein-positive was strongly influenced relative ratio to 2. In contrast, nearly all positive for Rep68/Rep40 were also expression. Examination...
ABSTRACT Adeno-associated virus type 2 (AAV-2) integrates specifically into a site on human chromosome 19 (chr-19) called AAVS1. To study the kinetics and frequency of chr-19-specific integration after AAV infection, we developed rapid, sensitive, quantitative real-time PCR assay for inverted terminal repeat-chr-19-specific junctions. Despite known variability junction sites, conditions were established that ensured reliable quantification rates within hours infection. The overall was...
Human Topors has originally been identified as binding partner of p53 and DNA topoisomerase I (TOP1). It can function both an ubiquitin SUMO‐1 E3 ligase for p53. Here we demonstrate that enhances the formation high‐molecular weight conjugates TOP1 in a reconstituted vitro system also human osteosarcoma cells, similar to treatment with CPT. In contrast situation observed p53, overall sumoylation levels were rather unaffected. Experiments point mutants strongly suggest represent chains formed...
Despite its very narrow tropism for erythroid progenitor cells, human parvovirus B19 (B19V) has recently been shown to replicate and form infectious progeny virus in 293 cells the presence of early adenoviral functions provided either by infection with adenovirus type 5 or addition pHelper plasmid encoding E2a, E4orf6, VA RNA functions. In present study we dissected individual influence these on B19V genome replication expression structural proteins VP1 VP2. We show that, constitutively...