Synthetische Nikotinamide in der Biokatalyse(S. 376)
Frank Hollmann, Caroline E. Paul
Natural nicotinamide cofactors (NAD(P)) can in some cases be replaced by simple synthetic analogues (mNADs). These analogues can enable ‘faster than naturally designed‘ enzymatic reactions, more efficient reaction processes and more chemoselective reactions because of their bioorthogonality.
Synthetische Biologie lässt Mikroorganismen „sehen“(S. 380)
Christian Renicke, Christof Taxis
Synthetic biology gave rise to microorganisms engineered to transform light cues to appropriate outputs to create images on living matter. Generation of such a image commonly requires enzymatic conversion of a colorless compound to a visible pigment. In yeast, this can be achieved by blocking the adenine biosynthesis pathway at a specific step: Fusing the respective enzyme to a photo-sensitive degron results in cells that allow photographic reproductions.
Analyse des „Ubiquitin-Codes“ mithilfe Protease-resistenter Ubiquitinketten(S. 382)
Tatjana Schneider, Daniel Schneider, Andreas Marx, Martin Schaffner
Modification of proteins by ubiquitin plays a fundamental role in maintaining a functional proteome. Proteins can be modified by single ubiquitin moieties or by various types of ubiquitin chains differing in linkage type and topology. Different ubiquitin chains are assumed to signal target proteins for different fates, but how this is achieved is only partially understood. We developed an easy-to-use strategy to generate large amounts of linkage-defined, non-hydrolyzable ubiquitin chains and show their potential to dissect ubiquitin signalling.
Proteintranslation und Prozessierung in physiologischer Umgebung abgebildet(S. 385)
Stefan Pfeffer, Förster Friedrich
Cryoelectron tomography allows 3D imaging of crowded pleiomorphic environments at molecular resolution and is consequently an excellent method for studying the structure and organization of large molecules in their natural context. Using this approach for the analysis of ribosomal complexes in different cellular compartments, we obtained detailed insights into the supramolecular organization of the cytosolic and mitochondrial translation machineries and their association to membranes for co-translational protein transport.
Organtypische Slice-Kulturen aus humanen Geweben(S. 388)
Felicitas Merz, Sonja Kallendrusch, Nikolas Schopow, Justus Körfer, Rasmus Sönnichsen
Preclinical research is often realized with cell lines or animal models and implies difficulties regarding the translation into clinics because of limitations of the models, e. g. species differences. We have therefore established methods to cultivate human tumor tissue from resections as organotypic slice cultures. Those slices contain all cells in their natural environment including the extracellular matrix. With this model, we aim to provide a test system which is closer to the situation in patients.
Dreidimensionale Vollhautrekonstrukte als Modelle humaner Erkrankungen(S. 392)
Dagmar Kulms, Silke Busch
Recently, three-dimensional skin reconstructs have proven to be very useful to investigate skinrelated diseases. In skin equivalents, keratinocytes, seeded on a dermal layer of fibroblasts embedded in a collagen scaffold, differentiate to form distinct layers of the epidermis as in natural skin. Above this, organotypic skin models can host melanoma nests or highly differentiated melanoma spheroids, making them suitable for investigating molecular targets for individualized melanoma treatment.
3D-Zellkultur zur Identifizierung von Zielmolekülen für die Krebstherapie(S. 397)
Ellen Dickreuter, Anne Vehlow, Nils Cordes
Malignant tumors often show radio- and chemoresistance phenotypes resulting from genetic mutations and epigenetic regulation. As molecular examinations for therapy optimization in patients and animal models are often challenging or infeasible, matrix-based 3-dimensional (3D) cell culture models have been developed as powerful tools in translational research to unravel the mechanisms of cancer cell survival, proliferation and metastasis as well as to identify potential cancer targets.
Designer-T-Zellen – neue Möglichkeiten für die Immuntherapie von Krebs(S. 402)
Matthias Leisegang, Wolfgang Uckert
The genetic engineering of T cells with T cell or chimeric antigen receptors generates tumor-specific designer T cells for immunotherapy of cancer. The clinical use of these cells requires (1) careful selection of the target antigen that should be tumor-specific, (2) an optimized configuration of therapeutic genes, to generate T cells of high functional activity and (3) efficient vector systems that allow the generation of sufficient numbers of engineered T cells within a short period of time.
Patientenspezifische iPS-Zellen und deren Anwendung in der Herzforschung(S. 406)
Katrin Streckfuß-Bömeke, Lukas Cyganek
A major research focus in the field of cardiovascular medicine is the prospect of using stem cells for cardiac regeneration. With the advent of induced pluripotent stem cell (iPSC) technology, major efforts are also in process to use iPSCs for modelling heart disease, screening for new drugs, and the development of treatment strategies on a patient-specific level. Here we describe the generation of iPSCs, their efficient differentiation into functional cardiomyocytes, and the power of the iPSC technology for modelling dilative cardiomyopathy on a patient-specific level.
Anwendungen & Produkte
Zytotoxizitäts-Assays mittels Imaging und Lumineszenz(S. 420)
Frauke Hänel, Norbert Garbow
Cytotoxicity is still a major cause for the failure of potential drugs in clinical trials and there is an urgent need for reliable and time-saving tests. Since compounds can interfere with a wide variety of biochemical processes, it is very important to analyze several parameters in ortho - gonal approaches.
On-line--pH- und -DO-Messungen in Mikrocarrier-basierter hMSC-Kultur(S. 422)
Valentin Jossen, Carmen Schirmaier, Gernot T. John, Dieter Eibl, Regine Eibl
Spinner flasks are often used for microcarrier-based cultivations of human mesenchymal stem cells (hMSCs). Normally, they are not equipped with pH and dissolved oxygen (DO) probes. This application note describes the cultivation of hMSCs in single-use spinner flasks equipped with optical pH (SP-HP8) and DO (SP-PSt3) sensors for the first time. While reaching peak cell numbers between 4.1 × 107 cells and 5.9 × 107 cells in two cultivation runs, reliable DO and pH data were delivered.
Biotechnologische Gewinnung von Seltenen Erden(S. 450)
Esther Gabor, Martin Langer, Jörg Reichert, Guido Meurer
Rare earth elements (REE) are ubiquitous on earth and several REEenriched deposits are known. However, their exploitation is technically and economically challenging. Recovery and raffination processes are highly complex and generally not very sustainable. In many high-tech products, however, REE constitute a crucial compound that can rarely sometimes be substituted by other elements. Novel biological approaches offer a promising option in the quest for safe, reliable, and economic REE winning processes.
Bioreactors go electro – Bioreaktoren für Bioelektrotechnologie aufrüsten(S. 453)
Carla Gimkiewicz, Steffi Hunger, Carolin Stang, Luis F. M. Rosa, Andreas Zehnsdorf, Falk Harnisch
Standardized bioreactors allowing systematic process engineering are established. The field of bioelectrotechnology, which combines the microbial or enzymatic conversion with electrical current flow, is still in its infancy. Thus, bioelectrochemical reactor systems are diverse, poorly characterized and not standardized. Here we present a reversible upgrade kit for existing bioreactor systems to perform bioelectrochemical syntheses – allowing a systematic process characterization and development.