Project Summary Core E - Virology, Next-Generation Sequencing and Imaging Core E is the main resource provided by the CFAR to support virological investigations of HIV/SIV and studies of its co-pathogens, such as Mycobacterium tuberculosis (Mtb) and Hepatitis C Virus (HCV). The Core has been configured to provide a wide range of services in support of modern virology research that now includes novel services for the rapid creation of recombinant viruses, production of recombinant virus and lentivirus vector stocks, support of shRNA library screens, high throughput, next-generation DNA sequencing (NGS). and access to high resolution fluorescence and electron microscopy technologies. The specific aims of the Virology, Next Generation Sequencing and Imaging core are: ? Virology: To provide access to panels of primary and recombinant HIV and SIV strains and recombinant yeast technology to create novel viral variants. The core maintains a repository of over 500 well- characterized primary HIV isolates as well as common and novel recombinant laboratory proviral constructs. Ongoing services include HIV, SIV, and lentiviral vector production and assay, provided on a per protocol basis. The core will also provide support for new technologies for shRNA library screens, which are in demand by numerous CFAR supported laboratories. ? Next Generation Sequencing (NGS): Provide access to the latest technologies in ultra-deep sequencing, including Ion Torrent, Roche 454, and Illumina platforms, in a CLIA/CAP (Clinical Laboratory Improvement Amendments/College of American Pathologists) accredited laboratory. In addition to offering the only deep sequencing-based HIV-1 Genotyping and Coreceptor Tropism Assay currently commercially available worldwide (DEEPGEN?HIV), the core has ample expertise and offers training in library preparation and assessment, sequencing, and bioinformatic analysis for both basic and clinical research projects. ? Imaging: Provide access and support for state-of-the-art fluorescence and electron microscopy across multiple platforms. The core houses a Deltavision fluorescence microscopy system within a biocontainment laboratory for live-cell experiments on HIV infected samples. The core supports other imaging platforms through strategic investments in Case imaging facilities to facilitate cross-campus collaborative interactions. In summary, Core E is provides invaluable support for HIV-1 projects involving molecular and cellular virology. The Core is continuously pushing forward new technologies that are not accessible to individual laboratories due to their high cost or degree of specialization. The Core will continue to extensive training and advice to its users and advance new technologies that fulfill unmet needs for the current and future HIV research priorities of the Case/UH CFAR.