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Lomonosov Moscow State University

5 ARTICLES PUBLISHED IN JoVE

Biology

Imaging Replicative Domains in Ultrastructurally Preserved Chromatin by Electron Tomography

Sophie Sosnovskaya^*1,2, Amir N. Zakirov^*1,2, Ekaterina D. Ryumina^*1,3, Ekaterina Kharybina^1,2, Sergei A. Golyshev¹, Olga S. Strelkova¹, Oxana A. Zhironkina¹, Andrei Moiseenko², Anton Orekhov⁴, Igor I. Kireev^1,2,5

¹Belozersky Institute of Physico-chemical Biology, Lomonosov Moscow State University, ²Faculty of Biology, Lomonosov Moscow State University, ³Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, ⁴National Research Center, Kurchatov Institute, ⁵V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology, and Perinatology

This protocol presents a technique for high-resolution mapping of replication sites in structurally preserved chromatin in situ that employs a combination of pre-embedding EdU-streptavidin-Nanogold labeling and ChromEMT.

Bioengineering

Microtubule Plus-End Dynamics Visualization in Huntington's Disease Model based on Human Primary Skin Fibroblasts

Aleksandra Taran^*1,2, Lilia Belikova (Shuvalova)^*2,3, Svetlana Lavrushkina^1,2, Alexandra Bogomazova^3,4, Maria Lagarkova^3,4, Irina Alieva^1,3

¹A.N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, ²Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, ³Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, ⁴Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency

This protocol is dedicated to the microtubule plus-end visualization by EB3 protein transfection to study their dynamic properties in primary cell culture. The protocol was implemented on human primary skin fibroblasts obtained from Huntington's disease patients.

Developmental Biology

Brain Organoid Generation from Induced Pluripotent Stem Cells in Home-Made Mini Bioreactors

Artem Eremeev^1,3, Lilia Belikova^1,2, Evgeny Ruchko¹, Egor Volovikov^1,5, Olga Zubkova¹, Alexy Emelin⁴, Roman Deev⁴, Olga Lebedeva^1,3, Alexandra Bogomazova^1,3, Maria Lagarkova^1,3

¹Federal Research and Clinical Center of Physical-Chemical Medicine, Federal Medical Biological Agency, ²Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, ³Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Federal Research and Clinical Center of Physical-Chemical Medicine, Federal Medical Biological Agency, ⁴Department of Pathological Anatomy, North-Western State Medical University, ⁵Department of Biology, Lomonosov State University

Here we describe a protocol for generating brain organoids from human induced pluripotent stem cells (iPSCs). To obtain brain organoids in large quantities and of high quality, we use home-made mini bioreactors.

Biochemistry

Analyzing the Interaction of Fluorescent-Labeled Proteins with Artificial Phospholipid Microvesicles using Quantitative Flow Cytometry

Nadezhda Podoplelova^1,2, Polina Soloveva^1,4, Andrei Garzon Dasgupta^1,2,3, Aleksandra Filkova^1,2, Mikhail Panteleev^1,2,3,4

¹Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Sciences, ²National Medical Research Center of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, ³Faculty of Physics, Lomonosov Moscow State University, ⁴Faculty of Biological and Medical Physics, Moscow Institute of Physics and Technology

Here, we describe a set of methods for characterizing the interaction of proteins with membranes of cells or microvesicles.

Biology

Exploring Caspase Mutations and Post-Translational Modification by Molecular Modeling Approaches

Dmitry K. Nilov¹, Alexey V. Zamaraev², Boris Zhivotovsky^2,3, Gelina S. Kopeina²

¹Belozersky Institute of Physicochemical Biology, Lomonosov Moscow State University, ²Faculty of Medicine, Lomonosov Moscow State University, ³Division of Toxicology, Institute of Environmental Medicine, Karolinska Institute

The present protocol uses a biomolecular simulation package and describes the molecular dynamics (MD) approach for modeling the wild-type caspase and its mutant forms. The MD method allows for assessing the dynamic evolution of the caspase structure and the potential effect of mutations or post-translational modifications.