Cell Biology

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    Optimized CRISPR-Cas9-based Strategy for Complex Gene Targeting in Murine Embryonic Stem Cells for Germline Transmission
    Authors:  Thomas J. O'Neill, Daniel Krappmann and Andreas Gewies, date: 05/20/2022, view: 149, Q&A: 0

    Although CRISPR-Cas9 genome editing can be performed directly in single-cell mouse zygotes, the targeting efficiency for more complex modifications such as the insertion of two loxP sites, multiple mutations in cis, or the precise insertion or

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    Preparation of a Single Cell Suspension from the Murine Iridocorneal Angle
    Authors:  Benjamin R. Thomson and Susan E. Quaggin, date: 05/20/2022, view: 40, Q&A: 0

    Single cell RNA sequencing is a powerful tool that can be used to identify distinct cell types and transcriptomic differences within complex tissues. It has proven to be especially useful in tissues of the eye, where investigators have identified

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    Preparation of Cas9 Ribonucleoproteins for Genome Editing
    Authors:  Sheng-Wei Lin, Viet Quoc Nguyen and Steven Lin, date: 05/20/2022, view: 70, Q&A: 0

    Genome editing by the delivery of pre-assembled Cas9 ribonucleoproteins (Cas9 RNP) is an increasingly popular approach for cell types that are difficult to manipulate genetically by the conventional plasmid and viral methods. Cas9 RNP editing is

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    CRISPR/Cas9 Gene Editing of HeLa Cells to Tag Proteins with mNeonGreen
    Authors:  Sachin Surve and Alexander Sorkin, date: 05/20/2022, view: 196, Q&A: 0

    Subcellular localization dynamics of proteins involved in signal transduction processes is crucial in determining the signaling outcome. However, there is very limited information about the localization of endogenous signaling proteins in living

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    Vestibular Organ Dissection and Whole-Mount Immunolabeling in Mouse
    Authors:  Timothy S. Balmer and Laurence O. Trussell, date: 05/20/2022, view: 56, Q&A: 0

    The vestibular sensory apparatus contained in the inner ear is a marvelous evolutionary adaptation for sensing movement in 3 dimensions and is essential for an animal’s sense of orientation in space, head movement, and balance. Damage to these

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    Protocol to Induce Follicular T Helper Cells, Germinal Centers, and Skin Lesions in Mouse Models for Skin Blistering Diseases

    Autoreactive T cells in autoantibody-mediated autoimmune diseases can be divided into two major subsets: (i) follicular T helper cells (Tfh) that provide T cell help in germinal centers (GC) and (ii) effector T (Teff) cells that immigrate into

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    A Flow Cytometry-Based Method for Analyzing DNA End Resection in G0- and G1-Phase Mammalian Cells
    Authors:  Bo-Ruei Chen, Jessica K. Tyler and Barry P. Sleckman, date: 05/20/2022, view: 82, Q&A: 0

    DNA double strand breaks (DSBs) constantly arise in cells during normal cellular processes or upon exposure to genotoxic agents, and are repaired mostly by homologous recombination (HR) and non-homologous end joining (NHEJ). One key determinant of

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    Protocol for Isolation of Cardiomyocyte from Adult Mouse and Rat
    Authors:  Huiliang Zhang and Peter S. Rabinovitch, date: 05/20/2022, view: 70, Q&A: 0

    The isolation of intact single adult cardiomyocytes from model animals, mouse and rat, is an essential tool for cardiac molecular and cellular research. While several methods are reported for adult mouse cardiomyocyte isolation, the viability and

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    Immunomagnetic Isolation and Enrichment of Microvascular Endothelial Cells from Human Adipose Tissue

    Human adipose tissue-resident microvascular endothelial cells are not only garnering attention for their emergent role in the pathogenesis of obesity-related metabolic disorders, but are also of considerable interest for vascular tissue engineering

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    Flow Cytometry-based Measurement of Reactive Oxygen Species in Cyanobacteria
    Authors:  Soumila Mondal and Shailendra P. Singh, date: 05/20/2022, view: 52, Q&A: 0

    Cyanobacteria are Gram-negative oxygen-producing photosynthetic bacteria that are useful in the pharmaceutical and biofuel industries. Monitoring of oxidative stress under fluctuating environmental conditions is important for determining the

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