Molecular Biology


    Protocols in Current Issue
    Production, Titration, Neutralisation, Storage and Lyophilisation of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Lentiviral Pseudotypes

    This protocol details a rapid and reliable method for the production and titration of high-titre viral pseudotype particles with the SARS-CoV-2 spike protein (and D614G or other variants of concern, VOC) on a lentiviral vector core, and use for neutralisation assays in target cells expressing angiotensin-converting enzyme 2 (ACE2) and

    Flow Cytometric Analysis of HIV-1 Transcriptional Activity in Response to shRNA Knockdown in A2 and A72 J-Lat Cell Lines
    Authors:  Daniela Boehm and Melanie Ott, date: 06/05/2017, view: 8171, Q&A: 0
    [Abstract] The main obstacle to eradicating HIV-1 from patients is post-integration latency (Finzi et al., 1999). Antiretroviral treatments target only actively replicating virus, while latent infections that have low or no transcriptional activity remain untreated (Sedaghat et al., 2007). To eliminate viral reservoirs, one strategy focuses ...
    Generation of Mutant Pigs by Direct Pronuclear Microinjection of CRISPR/Cas9 Plasmid Vectors
    Authors:  Chin-kai Chuang, Ching-Fu Tu and Chien-Hong Chen, date: 06/05/2017, view: 9918, Q&A: 0
    [Abstract] A set of Cas9 and single guide CRISPR RNA expression vectors was constructed. Only a very simple procedure was needed to prepare specific single-guide RNA expression vectors with high target accuracy. Since the de novo zygotic transcription had been detected in mouse embryo at the 1-cell stage, the plasmid DNA vectors encoding Cas9 and ...
    Murine Leukemia Virus (MLV)-based Coronavirus Spike-pseudotyped Particle Production and Infection
    Authors:  Jean Kaoru Millet and Gary R. Whittaker, date: 12/05/2016, view: 11926, Q&A: 0
    [Abstract] Viral pseudotyped particles (pp) are enveloped virus particles, typically derived from retroviruses or rhabdoviruses, that harbor heterologous envelope glycoproteins on their surface and a genome lacking essential genes. These synthetic viral particles are safer surrogates of native viruses and acquire the tropism and host entry pathway ...
    Reconstruction of the Mouse Inflammasome System in HEK293T Cells
    Authors:  Hexin Shi, Anne Murray and Bruce Beutler, date: 11/05/2016, view: 9410, Q&A: 0
    [Abstract] The NLRP3 (NLR family, Pyrin domain containing 3) inflammasome is a multiprotein complex comprised of NLRP3, pro-caspase-1, the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC), and the protein kinase NIMA related kinase 7 (NEK7) (Shi et al., 2016; He et al., 2016; Schmid-Burgk et al., ...
    Transfection of Human Naive CD4+ T Cells with PHA Activation and Neon Electroporation
    Authors:  Amy Palin and David B. Lewis, date: 08/05/2013, view: 15939, Q&A: 0
    [Abstract] Transfection of primary T cells can be challenging. This protocol describes a method to transfect primary human naive CD4+ T cells with an AP-1 luciferase reporter using low-level activation by phytohemagglutinin (PHA) and electroporation, as published (Palin et al., 2013). This technique is a modification of one previously ...
    Culture, Differentiation and Transfection of C2C12 Myoblasts
    Author:  Lili Jing, date: 05/20/2012, view: 51328, Q&A: 11
    [Abstract] C2C12 myoblasts are commonly used in biomedical laboratories as an in vitro system to study muscle development and differentiation. This protocol explains the basic procedures of culture, transfection and differentiation of C2C12 myoblast cells.

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