Core D-Get IN

Genetic modification, such as gene knockdown, overexpression, or precise genome editing, is the most effective way to elucidate gene function, which has the potential to unveil the mechanisms of skin diseases and advance therapeutic intervention. The delivery of genetic material into primary skin cells is notoriously difficult and delivery through the epidermal barrier is a great challenge. The Gene Editing, Transduction, and Nanotechnology (GET iN) Core will apply the most advanced and innovative approaches for genetic manipulation. These include CRISPR/Cas9 technologies and both viral and non-viral methods for the delivery of DNA, RNA, and other biomolecules into skin cells in 2D and 3D cultures, skin explants, and intact skin. Specifically, the Core will generate highly consistent viral stocks in a broad range of systems (lenti/retro/adeno/AAV) for gene manipulation using siRNA, cDNA, and Cas9/sgRNA. The Core will also facilitate the use of spherically oriented nanocarriers (e.g., spherical nucleic acids and high-density lipoprotein nanoparticles) for intracellular and transcutaneous DNA, RNA, protein and drug delivery to skin cells and skin. The Core will provide SBDRC members with equipment and necessary reagents for nucleoporation. Services related to CRISPR include CRISPR/Cas9- mediated gene knockout, reversible gene expression manipulation via epigenomic approaches (CRISPRa and CRISPRi), and genome-wide CRISPR screen capabilities, widely used for functional gene discovery. As another new service we will also provide reagents and offer expert advice on novel approaches for genome editing, such as single base editing. The GET iN Core will maintain human and mouse lentiviral shRNA libraries, establish a panel of frequently used reagents (viral vectors, DNA/RNA inserts, CRISPR gRNAs, and genetically modified skin cells) for skin research, and offer reagents for CRISPR technology. The Core will sponsor educational and consultation programs, given by both internal and external speakers, on the topics of: effective delivery of genetic material, gene manipulation using nanotechnology, CRISPR technology, genome editing, genetic screens, and gene therapy. Having a wide range of customized viral vectors and unique reagents, novel CRISPR/Cas9 services, and innovative nanotechnology approaches, this Core will provide the most advanced tactics for genetic manipulation in 2D and 3D skin cultures, as well as skin in vivo. Our services, and collaboration with other SBDRC and Northwestern Cores, will further enhance skin biology and diseases research at NU and nationally, ultimately facilitating the bench to bedside translation of research.