Super micrometer, measuring machine, gage, measurement, micrometer. Keywords, supermic, labmaster, metrology, calibration, supermicrometer, measuring. These super-powered lines can contain as many line segments as you want, and.The immature virion consists of approximately 2500 Gag molecules arranged as spherical protein shell. The virus is initially released as immature non-infectious particle consisting of mostly uncleaved Gag polyproteins. CRISP/Cas based approaches for HIV‑1 eliminationFigure 1: Tomogram of HIV particles (red) budding at the plasma membrane (blue)Assembly and budding of Human Immunodeficiency Virus (HIV) is directed by the main structural polyprotein Gag and occurs at the plasma membrane of the infected cell.These provide a better understanding of the mechanisms and protein interfaces governing assembly and maturation.Figure 2: Post-entry steps in retroviral replication. These studies are complemented by analyses of HIV particle formation in tissue culture.We employ structural analyses including transmission EM, cryo-EM and tomography to obtain detailed three-dimensional images of mature and immature virions, as well as viral budding sites, of wild-type and mutant HIV. We have developed in vitro assembly systems to analyze the molecular architecture of capsid assemblies, to study the effect of mutations on mature and immature particle formation and identify inhibitors of HIV assembly. Maturation leads to disassembly of the immature Gag layer, followed by a second assembly stage forming the mature cone-shaped capsid that incorporates the condensed viral genome associated with replication proteins.The assembly of immature particles requires only the viral Gag polyprotein and can be mimicked in vitro using bacterially purified Gag-derived proteins.These early post-entry replication steps occur within poorly charaterized subviral complexes termed reverse-transcription and pre-integration complexes (RTC/PIC). We are particularly interested in (i) cGAS, a component of the innate immune system that detects the presence of viral DNA, (ii) TREX1 digests abortive viral cDNA thus downregulating the immune response to MLV and HIV-1 infection, (iii) NONO, that binds directly to capsid and is essential for cGAS activation and relocalization to the nucleus.Delivery of the genome-containing retroviral capsid into the cytoplasm of a newly infected cell is followed by reverse transcription of the viral RNA and transport of the resulting cDNA to the host cell DNA, where the genetic information of the virus is integrated. Numerous host cell factors can target viral replication complexes. The capsid has to navigate through a hostile cytoplasmic environment while the RNA genome is reverse transcribed into DNA.
![]() While many retroviruses (e.g. Finally, there are fundamental differences not only between different types of host cells, but also between different retroviruses. Furthermore, numerous host cell dependency and restricition factors affect these processes in a cell type dependent manner. Dabster anime one piece episode 435 reactioninvestigate the function of host cell dependency (e.g. characterize the composition and structure of retroviral replication complexes in different cells and at different intracellular localizations, HIV-1, can enter the intact nucleus through nuclear pores.Our work on retroviral post-entry comprises a number of different projects, in which we History google chromeIn these projects, we collaborate with different groups within the Collaborative Research Center SFB 1129 and the Priority Program SPP 1923, funded by the German Research Foundation (DFG).HIV is an enveloped virus. Digital droplet PCR), virological and biochemical approaches. EdU-click labeling of viral DNA, 3D FISH ANCHOR visualization) with state-of-the-art microscopy (STED nanoscopy, correlative light and electron microscopy, (cryo)electron tomography, live cell imaging), molecular biological (e.g. and compare replication pathways and cellular immune responses in HIV-1 and MLV infected cells.For this, we combine advanced labeling technologies (e.g. assess the role of capsid stability in post-entry processes CGAS, TREX1) that detect the viral cDNA, Using clickable lipid analogs and STED nanoscopy we investigate the (re)distribution of distinct lipids during the viral assembly process.Current anti-retroviral therapy (cART) can prevent destruction of the immune system of HIV-1 infected patients and protects from developing AIDS disease. We are currently analyzing the influence of host cells and viral proteins on the lipid compostion of the envelope and study the influence of lipid composition on the properties of the virus.In order to study the lipid and protein organization at HIV-1 assembly sites, we have developed a system that allows us to recruit the viral structural protein Gag to the plasma membrane at a defined time point. However, unlike viral proteins, viral lipids and their functional role a poorly understood.In collaboration with Britta Brügger we have determined the lipidome of HIV using mass spectroscopy and biochemical approaches. Thus, lipids are involved in virus entry and egress, and they may play functional roles in viral protein composition, particle stability and infectivity. During infection of new host cells, this lipid envelope fuses to the cellular plasma membrane. By directing the machinery to the viral genome we predict that we can inactivate the pathogenic sequences, thereby curing the infected cells (and, eventually, patients). CRISPR/Cas can be targeted to cleave and modify a desired DNA sequence. In collaboration with the group of Dirk Grimm ( Grimm Lab), we develop T-cell specific viral gene transfer vectors and use them to express components of the CRISPR/Cas system in HIV-infected cells. Interruption of therapy thus results in renewed virus production from latent reservoirs.Within this project we aim at fully eradicating HIV by combining state-of-the art gene delivery and genome engineering technologies. HIV-1 integrates its genetic information into the host genome and can thus not be eliminated from the organism once the infection is established. How these filamentous particles are formed and how they are taken up into the next host cell is currently not understood.For our studies we develop fluorescently labeled influenza virus derivatives that allow us to analyze replication and spread of spheroidal and filamentous particles by live cell imaging and superresolution microscopy. While most studies to date were carried out using lab adapted influenza variants that form small spherical particles, most influenza viruses from patient samples appear to be micrometer long filaments. Our group wants to obtain a better understanding of the formation and spread of influenza virus. Despite intense research efforts, currently available vaccines and virostatics have significant limitations. Cell nuclei (DraQ5) and the mucin 1 transmembrane protein are stained for visualization.Influenza virus is a major human pathogen that affects roughly 10% of the world population each year. They comprise a variety of specialized cells typical for a specific organ and thus mimick the complex environment that the virus encounters in vivo. Organoids are millimeter sized three-dimensional microstructures which can be grown from various types of stem cells.
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