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Helmholtz Zentrum München

3 ARTICLES PUBLISHED IN JoVE

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Genetics

Real-time Analysis of Transcription Factor Binding, Transcription, Translation, and Turnover to Display Global Events During Cellular Activation
Kathrin Davari *1, Johannes Lichti *1, Caroline C. Friedel 2, Elke Glasmacher 1,3
1Institute for Diabetes and Obesity (IDO), German Center for Diabetes Research (DZD), Helmholtz Zentrum München, 2Institute for Informatics, Ludwig-Maximilians-Universität München, 3Roche Pharma Research and Early Development, Large Molecule Research, Roche Innovation Center Penzberg

This protocol describes the combinatorial use of ChIP-seq, 4sU-seq, total RNA-seq, and ribosome profiling for cell lines and primary cells. It enables tracking changes in transcription-factor binding, de novo transcription, RNA processing, turnover and translation over time, and displaying the overall course of events in activated and/or rapidly changing cells.

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Genetics

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)
Christopher T. Breunig 1,2, Andrea M. Neuner 1,2, Jessica Giehrl-Schwab 3, Wolfgang Wurst 3, Magdalena Götz 2,4, Stefan H. Stricker 1,2
1MCN Junior Research Group, Munich Center for Neurosciences, Ludwig Maximilian Universitat, BioMedical Center, 2Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, 3Institute of Developmental Genetics, Helmholtz Zentrum, German Research Center for Environmental Health, 4Physiological Genomics, Ludwig Maximilian Universitat, BioMedical Center

Here, we present string assembly gRNA cloning (STAgR), a method to easily multiplex gRNA vectors for CRISPR/Cas9 approaches. STAgR makes gRNA multiplexing simple, efficient and customizable.

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Neuroscience

Cryo-section Dissection of the Adult Subependymal Zone for Accurate and Deep Quantitative Proteome Analysis
Christian Friess 1, Magdalena Götz 1,2,3, Jacob Kjell 1,2,4
1Division of Physiological Genomics, Biomedical Center, Ludwig Maximilian University of Munich, 2Institute for Stem Cell Research, Helmholtz Zentrum München, 3SYNERGY, Excellence Cluster Systems Neurology, University of Munich, 4Department of Clinical Neuroscience, Karolinska Institutet

Cryo-section-dissection allows fresh, frozen preparation of the largest neurogenic niche in the murine brain for deep quantitative proteome analysis. The method is precise, efficient, and causes minimal tissue perturbation. Therefore, it is ideally suited for studying the molecular microenvironment of this niche, as well as other organs, regions, and species.

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