Protocols in Current Issue
    Unbiased Screening of Activated Receptor Tyrosine Kinases (RTKs) in Tumor Extracts Using a Mouse Phospho-RTK Array Kit
    Authors:  Julian Naipauer, Lucas E. Cavallin and Enrique A. Mesri, date: 04/20/2019, view: 381, Q&A: 0
    [Abstract] Kaposi’s sarcoma (KS) herpesvirus (KSHV) is a virus that causes KS, an angiogenic AIDS-associated spindle-cell neoplasm, by activating host oncogenic signaling cascades through autocrine and paracrine mechanisms. Many host signaling cascades co-opted by KSHV including PI3K/AKT/mTORC, NFkB and Notch are critical for cell-specific mechanisms of ...
    Quantifying Symmetrically Methylated H4R3 on the Kaposi’s Sarcoma-associated Herpesvirus (KSHV) Genome by ChIP-Seq
    Authors:  Roxanne C. Strahan, Kayla S. Hiura and Subhash C. Verma, date: 03/20/2018, view: 2348, Q&A: 0
    [Abstract] Post-translational modifications to histone tails contribute to the three-dimensional structure of chromatin and play an important role in determining the relative expression of nearby genes. One such modification is symmetric di-methylation of arginine residues, which may exhibit different effects on gene expression including blocking the binding ...
    A Bioreactor Method to Generate High-titer, Genetically Stable, Clinical-isolate Human Cytomegalovirus
    [Abstract] Human cytomegalovirus (HCMV) infection is a major cause of morbidity and mortality in transplant patients and a leading cause of congenital birth defects (Saint Louis, 2016). Vaccination and therapeutic studies often require scalable cell culture production of wild type virus, represented by clinical isolates. Obtaining sufficient stocks of ...
    Vaginal HSV-2 Infection and Tissue Analysis
    Authors:  Marie Beck Iversen, Søren Riis Paludan and Christian Kanstrup Holm, date: 07/05/2017, view: 4246, Q&A: 0
    [Abstract] The vaginal murine HSV-2 infection model is very useful in studying mucosal immunity against HSV-2 (Overall et al., 1975; Renis et al., 1976; Parr and Parr, 2003). Histologically, the vagina of Depo-Provera-treated mice is lined by a single layer of mucus secreting columnar epithelial cells overlying two to three layers of ...
    Transient Transfection-based Fusion Assay for Viral Proteins
    Authors:  Melina Vallbracht, Christina Schröter, Barbara G. Klupp and Thomas C. Mettenleiter, date: 03/05/2017, view: 5142, Q&A: 1
    [Abstract] Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) glycoproteins gB, gH and gL. Here, we describe an ...
    ACE-score-based Analysis of Temporal miRNA Targetomes During Human Cytomegalovirus Infection Using AGO-CLIP-seq
    Authors:  Sungchul Kim and Kwangseog Ahn, date: 04/20/2016, view: 5677, Q&A: 0
    [Abstract] Although temporal regulation of gene expression during the course of infection is known to be critical for determining the outcome of host-virus interactions, systematic temporal analysis of the miRNA targetomes during productive viral infection has been technically challenging due to the large range of miRNA-mRNA cross-talks at the host-virus ...
    Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses
    Authors:  Anna Sloutskin and Ronald S. Goldstein, date: 11/20/2014, view: 25605, Q&A: 0
    [Abstract] Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for fluorescently labeled viruses, based on the original plaque assay. In addition, the calculation of multiplicity of infection (MOI) is presented.
    siRNA Screening for Genes Involved in HSV-1 Replication
    Authors:  Samantha J. Griffiths and Jürgen Haas, date: 08/20/2014, view: 6520, Q&A: 0
    [Abstract] Small interfering RNAs (siRNAs) are small (typically 18-24 nucleotides) RNA molecules capable of silencing gene expression post-transcriptionally and as such, they provide a simple method by which the role of individual genes in complex cellular systems can be easily assessed. As siRNAs are easy to use experimentally, and can be designed to target ...
    VZV Replication Assays
    Authors:  Samantha J. Griffiths and Jürgen Haas, date: 08/20/2014, view: 6939, Q&A: 0
    [Abstract] Varicella zoster virus (VZV) is a human herpesvirus which causes Varicella (chickenpox) upon primary infection and Zoster (shingles) following reactivation from latency (von Bokay, 1909). Whilst VZV is extensively studied, inherent features of VZV replication, such as cell-association of virus particles during in vitro culture and a ...
    Purification of Herpesvirus Virions and Capsids
    Authors:  Xinghong Dai and Z. Hong Zhou, date: 08/05/2014, view: 9398, Q&A: 4
    [Abstract] This protocol was designed for large-scale purification of herpesvirus particles by cell culture. Virions and capsids are isolated from extracellular culture media and cell nuclei, respectively. Purity and concentration of the purified samples are usually sufficient for structural studies with cryo electron microscopy and cryo electron tomography. ...

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