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Virology

protocols

Protocol

Protocol for Delivering AAV to Express Fluorescent Biosensors in the Juvenile Mouse Hippocampus

Experiment Tags: Adeno-Associated Viruses, Acute Brain Slices, Genetically-Encoded Fluorescent Biosensors, Hippocampus, Intracranial Injection, Dentate Granule cells

Experiment Summary: In this protocol, we describe a protocol for intracranial injections of two viral vectors encoding the metabolic biosensor Peredox and the calcium biosensor RCaMP1h. When combined with 2-photon microscopy and fluorescence lifetime imaging, this protocol allows the simultaneous quantitative assessment of changes in the cytosolic NADH/NAD+ ratio and the intracellular Ca2+ levels in individual dentate granule cells from acute hippocampal slices.

protocols

Protocol

Protocol for Intrathoracic Inoculation of Zika Virus in Aedes aegypti

Experiment Tags: Aedes aegypti, Mosquito, Zika virus, Arbovirus, Intrathoracic Inoculation, Microinjection

Experiment Summary: This protocol describes herein a protocol for the intrathoracic inoculation of ZIKV in Ae. aegypti. This method bypasses the midgut, which leads to a more rapid and higher proportion of disseminated infections in comparison to oral challenge, and mosquitoes become infected with a consistent dose of virus.

protocols

Protocol

Varicella Zoster Virus Replication Assays

Experiment Tags: Varicella Zoster Virus (VZV), Replication Assays

Experiment Summary: This protocol describes a basic VZV replication assay using a recombinant VZV-GFP reporter virus. This reporter virus-infected cell line can be used in combination with siRNA gene depletion or cDNA overexpression transfection protocols to determine the effect of individual cellular genes on virus replication.

protocols

Protocol

Antibody-Based CAR T Cells Produced by Lentiviral Transduction Protocol

Experiment Tags: Lentiviral Transduction, CAR T cells

Experiment Summary: In this protocol, an easy and reliable approach for CAR T cell generation is described. T cells are first isolated from peripheral blood and afterwards activated for one day with anti-CD3/CD28 Dynabeads. The gene transfer is performed by lentiviral transduction and gene transfer rate can be verified by flowcytometric analysis.

protocols

Protocol

Retroviral Capsid Core Stability Assay Protocol

Experiment Tags: Retrovirus, Enveloped Virus, MLV, HIV, Capsid Core Stability Assay, Fate of Capsid Assay, Glycosylated Gag, gPr80, TRIM5α

Experiment Summary: In this protocol, we describe a method using an in vitro detergent-based approach combined with ultracentrifugation for assessing viral capsid core stability. The results show that this method is a significantly simpler and faster way to assess relative capsid core stability.

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