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Background Plasma concentration of low-density lipoprotein (LDL) cholesterol is a well-established risk factor for cardiovascular disease. Inhibition of proprotein convertase subtilisin/kexin type 9 (PCSK9), which regulates cholesterol homeostasis, has recently emerged as an approach to reduce cholesterol levels. The development of humanized animal models is an important step to validate and study human drug targets, and use of genome and base editing has been proposed as a mean to target disease alleles . Results To address the lack of validated models to test the safety and efficacy of techniques to target human PCSK9, we generated a liver-specific human PCSK9 knock-in mouse model (hPCSK9-KI). We showed that plasma concentrations of total cholesterol were higher in hPCSK9-KI than in wildtype mice and increased with age. Treatment with evolocumab, a monoclonal antibody that targets human PCSK9, reduced cholesterol levels in hPCSK9-KI but not in wildtype mice, showing that the hypercholesterolemic phenotype was driven by overexpression of human PCSK9. CRISPR-Cas9-mediated genome editing of human PCSK9 reduced plasma levels of human and not mouse PCSK9, and in parallel reduced plasma concentrations of total cholesterol; genome editing of mouse Pcsk9 did not reduce cholesterol levels. Base editing using a guide RNA that targeted human and mouse PCSK9 reduced plasma levels of human and mouse PCSK9 and total cholesterol. In our mouse model, base editing was more precise than genome editing, and no off-target editing nor chromosomal translocations were identified. Conclusions Here, we describe a humanized mouse model with liver-specific expression of human PCSK9 and a human-like hypercholesterolemia phenotype, and demonstrate that this mouse can be used to evaluate antibody and gene editing-based (genome and base editing) therapies to modulate the expression of human PCSK9 and reduce cholesterol levels. We predict that this mouse model will be used in the future to understand the efficacy and safety of novel therapeutic approaches for hypercholesterolemia.

The CRISPR-Cas9 system has increased the speed and precision of genetic editing in cells and animals. However, model generation for drug development is still expensive and time-consuming, demanding more target flexibility and faster turnaround times with high reproducibility. The generation of a tightly controlled ObLiGaRe doxycycline inducible SpCas9 (ODInCas9) transgene and its use in targeted ObLiGaRe results in functional integration into both human and mouse cells culminating in the generation of the ODInCas9 mouse. Genomic editing can be performed in cells of various tissue origins without any detectable gene editing in the absence of doxycycline. Somatic in vivo editing can model non-small cell lung cancer (NSCLC) adenocarcinomas, enabling treatment studies to validate the efficacy of candidate drugs. The ODInCas9 mouse allows robust and tunable genome editing granting flexibility, speed and uniformity at less cost, leading to high throughput and practical preclinical in vivo therapeutic testing. CRISPR/Cas9 technology has revolutionised the ability of scientists to make genetically modified cells and animal models. Here, the authors make a Tet-On genetically modified mouse using the Streptococcus pyogenes Cas9 and demonstrate that it can be used for generation of mice with lung cancer.

The paracaspase mucosa-associated lymphoid tissue lymphoma translocation protein-1 (MALT1) regulates nuclear-factor-kappa-B (NF-[kappa]B) activation downstream of surface receptors with immunoreceptor tyrosine-based activation motifs (ITAMs), such as the B-cell or T-cell receptor and has thus emerged as a therapeutic target for autoimmune diseases. However, recent reports demonstrate the development of lethal autoimmune inflammation due to the excessive production of interferon gamma (IFN-É£) and defective differentiation of regulatory T-cells in genetically modified mice deficient in MALT1 paracaspase activity. To address this issue, we explored the effects of pharmacological MALT1 inhibition on the balance between T-effector and regulatory T-cells. Here we demonstrate that allosteric inhibition of MALT1 suppressed Th1, Th17 and Th1/Th17 effector responses, and inhibited T-cell dependent B-cell proliferation and antibody production. Allosteric MALT1 inhibition did not interfere with the suppressive function of human T-regulatory cells, although it impaired de novo differentiation of regulatory T-cells from naïve T-cells. Treatment with an allosteric MALT1 inhibitor alleviated the cytokine storm, including IFN-É£, in a mouse model of acute T-cell activation, and long-term treatment did not lead to an increase in IFN-É£ producing CD4 cells or tissue inflammation. Together, our data demonstrate that the effects of allosteric inhibition of MALT1 differ from those seen in mice with proteolytically inactive MALT1, and thus we believe that MALT1 is a viable target for B and T-cell driven autoimmune diseases.

Osteogenesis imperfecta (OI) is a hereditary disease occurring in humans and dogs. It is characterized by extremely fragile bones and teeth. Most human and some canine OI cases are caused by mutations in the COL1A1 and COL1A2 genes encoding the subunits of collagen I. Recently, mutations in the CRTAP and LEPRE1 genes were found to cause some rare forms of human OI. Many OI cases exist where the causative mutation has not yet been found. We investigated Dachshunds with an autosomal recessive form of OI. Genotyping only five affected dogs on the 50 k canine SNP chip allowed us to localize the causative mutation to a 5.82 Mb interval on chromosome 21 by homozygosity mapping. Haplotype analysis of five additional carriers narrowed the interval further down to 4.74 Mb. The SERPINH1 gene is located within this interval and encodes an essential chaperone involved in the correct folding of the collagen triple helix. Therefore, we considered SERPINH1 a positional and functional candidate gene and performed mutation analysis in affected and control Dachshunds. A missense mutation (c.977C>T, p.L326P) located in an evolutionary conserved domain was perfectly associated with the OI phenotype. We thus have identified a candidate causative mutation for OI in Dachshunds and identified a fifth OI gene.

Im September fand das 10. Wildauer Bibliothekssymposium statt und stellte sich in ganzer thematischer Breite dem Angebotsspektrum Innovation bzw. womit sich Bibliotheken aktuell dazu beschäftigen. Die Palette reicht von Stegreiftheater bis institutioneller Neuaufstellung, vom Einsatz humanoider Roboter bis zur Anwendung von Algorithmen des maschinellen Lernens für Suchanfragen.

CRISPR–Cas genome-editing nucleases hold substantial promise for developing human therapeutic applications 1 – 6 but identifying unwanted off-target mutations is important for clinical translation 7 . A well-validated method that can reliably identify off-targets in vivo has not been described to date, which means it is currently unclear whether and how frequently these mutations occur. Here we describe ‘verification of in vivo off-targets’ (VIVO), a highly sensitive strategy that can robustly identify the genome-wide off-target effects of CRISPR–Cas nucleases in vivo. We use VIVO and a guide RNA deliberately designed to be promiscuous to show that CRISPR–Cas nucleases can induce substantial off-target mutations in mouse livers in vivo. More importantly, we also use VIVO to show that appropriately designed guide RNAs can direct efficient in vivo editing in mouse livers with no detectable off-target mutations. VIVO provides a general strategy for defining and quantifying the off-target effects of gene-editing nucleases in whole organisms, thereby providing a blueprint to foster the development of therapeutic strategies that use in vivo gene editing. A strategy developed to define off-target effects of gene-editing nucleases in whole organisms is validated and leveraged to show that CRISPR–Cas9 nucleases can be used effectively in vivo without inducing detectable off-target mutations.

Innovationen vielfältigster Art auf Informationseinrichtungen heruntergebrochen, dafür stand abermals das letzte, achte Wildauer Bibliothekssymposium. D. h. diskutiert wurden Innovation als agile Projektmanagementmethode, strategisches Programm, unveräußerliche Kernaufgabe von Bibliotheksteams, als Chance für unbemannte Services (Öffnungszeiten), aber auch, was dies für die technischen Kompetenzen bedeutet. Nach Rezeption aller Vorträge, die auch Themen wie chaotische Lagerhaltung, RFID-Inventur, Serendipität tangierten, bleibt das Fazit, jede Bibliothek birgt Unmengen von Entwicklungspotentialen, die ihren Deut beitragen, der Intention von Bibliotheken ihren Zielgruppen gegenüber zeitgemäß gerechter zu werden

Die Wildauer Hochschulbibliothek hatte sich 2013 trotz guter „Bilanzen“ entschlossen, wegen der auf sie kommenden Unwägbarkeiten und Herausforderungen ein strategisches wie machbares Konzept zu erarbeiten. Dieses soll bis zum Jahr 2020 umgesetzt sein und die Entscheider im Umfeld der Hochschulbibliothek über ihre Ziele und Meilensteine verständigen. Die Stakeholder sollen über die schriftliche Fixierung ebenfalls dafür gewonnen und begeistert werden. Das ambitionierte Werk, die Agenda 2020, startete damit, dass das Bibliotheksteam fern vom Alltag an einem ungewöhnlichen, aber inspirierenden Ort in Klausur ging. Ein Segeltörn zu Pfingsten 2013 gab den Start für die Agenda 2020. Sieben Themenfelder konnten identifiziert und mit konkreten Vorhaben formuliert werden. Despite its good standing the university library of Wildau decided last year, to develop a strategic and viable concept to face future challenges and uncertainties. The concept will cover the period up until the year 2020. It is supposed to provide transparency by informing the decision-makers of the university about planned objectives, goals and milestones. Besides, the support of stakeholders is to be won by providing them with a written strategy paper. The work on the Agenda 2020 started with an event outside the campus and library. As an unusual but inspiring place to retreat the library team chose a sailing boat. A cruise during Pentecost 2013 announced the launch of the Agenda 2020. Seven topics were identified and formulated including specific projects.