Molecular Biology

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    Genome-wide Mapping of 5′-monophosphorylated Ends of Mammalian Nascent RNA Transcripts
    Authors:  Michael A. Cortázar, Nova Fong and David L. Bentley, date: 09/20/2023, view: 83, Q&A: 0

    In eukaryotic cells, RNA biogenesis generally requires processing of the nascent transcript as it is being synthesized by RNA polymerase. These processing events include endonucleolytic cleavage, exonucleolytic trimming, and splicing of the growing

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    Isolation of Epithelial and Stromal Cells from Colon Tissues in Homeostasis and Under Inflammatory Conditions

    Inflammation of the gastrointestinal tract is a prevalent pathology in diseases such as inflammatory bowel disease (IBD). Currently, there are no therapies to prevent IBD, and available therapies to treat IBD are often sub-optimal. Thus, an unmet

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    Fluorescence in situ Localization of Pri-miRNAs in Isolated Arabidopsis thaliana Nuclei

    Here, we present an approach combining fluorescence in situ hybridization (FISH) and immunolabeling for localization of pri-miRNAs in isolated nuclei of A. thaliana. The presented method utilizes specific DNA oligonucleotide probes, modified by

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    Successful Transfection of MicroRNA Mimics or Inhibitors in a Regular Cell Line and in Primary Cells Derived from Patients with Rheumatoid Arthritis
    Authors:  Si Wang, Jing Xu, Yuanxu Guo, Yongsong Cai, Wenhua Zhu, Liesu Meng, Congshan Jiang and Shemin Lu, date: 09/20/2023, view: 66, Q&A: 0

    The transfection of microRNA (miRNA) mimics and inhibitors can lead to the gain and loss of intracellular miRNA function, helping us better understand the role of miRNA during gene expression regulation under specific physical conditions. Our

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    Fractionation of Native Protein Complexes from Mammalian Cells to Determine the Differential Proteasome Activity and Abundance
    Authors:  Ana Paula Zen Petisco Fiore and Christine Vogel, date: 09/20/2023, view: 158, Q&A: 0

    Eukaryotic cells have different types of proteasomes that differ in size. The smallest proteolytically active particle is the 20S proteasome, which degrades damaged and oxidized proteins; the most common larger particle is the 26S proteasome, which

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    Immunoprecipitation of Reporter Nascent Chains from Active Ribosomes to Study Translation Efficiency
    Authors:  Roberta Cacioppo and Catherine Lindon, date: 09/20/2023, view: 91, Q&A: 0

    The study of translation is important to the understanding of gene expression. While genome-wide measurements of translation efficiency (TE) rely upon ribosome profiling, classical approaches to address translation of individual genes of interest

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    Controlled Level of Contamination Coupled to Deep Sequencing (CoLoC-seq) Probes the Global Localisation Topology of Organelle Transcriptomes
    Authors:  Anna Smirnova, Damien Jeandard and Alexandre Smirnov, date: 09/20/2023, view: 131, Q&A: 0

    Information on RNA localisation is essential for understanding physiological and pathological processes, such as gene expression, cell reprogramming, host–pathogen interactions, and signalling pathways involving RNA transactions at the level

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    A Post-translational Modification–enhanced Pull-down Method to Study Degron Domains and the Associated Protein Degradation Complexes
    Authors:  Pierluigi Scalia and Stephen J. Williams, date: 09/20/2023, view: 183, Q&A: 0

    The identification and characterization of the ubiquitin E-ligase complexes involved in specific proteins’ degradation via the ubiquitin-proteasome system (UPS) can be challenging and require biochemical purification processes and in vitro

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    Spatial Centrosome Proteomic Profiling of Human iPSC-derived Neural Cells
    Authors:  Fatma Uzbas and Adam C. O’Neill, date: 09/05/2023, view: 213, Q&A: 0

    The centrosome governs many pan-cellular processes including cell division, migration, and cilium formation. However, very little is known about its cell type-specific protein composition and the sub-organellar domains where these protein

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    Absolute Quantification of mRNA Isoforms in Adult Stem Cells Using Microfluidic Digital PCR
    Authors:  Shubhangi Das Barman, Zofija Frimand and Antoine de Morree, date: 09/05/2023, view: 117, Q&A: 0

    Adult stem cells play key roles in homeostasis and tissue repair. These cells are regulated by a tight control of transcriptional programs. For example, muscle stem cells (MuSCs), located beneath the basal lamina, exist in the quiescent state but

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