Gene Editing and Virus Production Module

Resource/Service Core: Gene Editing and Virus Production Module Abstract The Gene Editing and Virus Production (GEVP) Module will provide access to state-of-the-art genome editing technologies and generate DNA/RNA constructs for use in mammalian cultures/systems in vision research. This Module will also purify and concentrate both lentivirus and adeno-associated virus (AAV) for subsequent use in immortalized, primary cultures (maintained by individual investigators and the Stem Cell, Organoid and Cell Phenotyping Module) and in animals in vivo. The specific services provided within the GEVP Module are; 1) CRISPR guide-RNA to perform DNA editing or epigenetic modifications, 2) site-directed mutagenesis for generating point mutations, insertions, or deletion variants, 3) shRNA plasmid generation for knockdown/out experiments, 4) full sequencing of generated constructs, extensive documentation, and maintenance of DNA stocks for future use, 5) small to large scale endotoxin free plasmid purification for downstream applications, thorough confirmation of produced constructs at the transcript, protein, and subcellular level, consultation for optimal promoters and serotypes, and 6) generation of high-purity viruses for use in vitro or in vivo. Overall, the GEVP Module will provide access to leading-edge genome editing technologies, support a range of DNA/RNA and viral technologies, and provide the assistance and expertise required to test unique hypotheses posited by vision scientists at UTSW and neighboring institutions.