Transgenic & Gene-Targeting Shared Resource

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TRANSGENIC AND GENE-TARGETING SHARED RESOURCE (TGTSR): ABSTRACT Genetically altered models are important tools for the researchers at The University of Kansas Cancer Center (KUCC). The production and analysis of such models using CRISPR-based genome editing methods, including mouse, pluripotent stem cell, and tumor cell line models, ultimately leads to a better understanding of the nature, progression, and functional genomics of tumor formation. Genetically engineered mouse strains also serve as in vivo models for diagnostics and treatment. The techniques employed to generate genetically modified animal and cell models require specialized equipment and technical expertise. The Transgenic and Gene-Targeting Shared Resource (TGTSR) led by Jay L. Vivian, PhD, supports members of KUCC by providing centralized and comprehensive technical services for the production of novel genetically engineered rodents and cell lines. The TGTSR uses cutting-edge methods, state-of-the-art instrumentation, and novel reagents for the generation of these models. Genome editing methods are central to the activities of the TGTSR, including a pipeline for the design and optimized use of CRISPR reagents in embryo and cell models. The expertise of the TGTSR Director and staff is leveraged in all phases of the generation of novel genetically engineered models, from the initial experimental design stage through model generation, molecular characterization, expansion, genotyping, and cryopreservation. This extensively used shared resource supported 28 KUCC users in CY2020, and 49 unique KUCC members were supported by the TGTSR in the previous funding period, demonstrating the broad use of this shared resource. The five full-time staff members of the TGTSR include the coordinated efforts between staff at both the University of Kansas Medical Center (KUMC) and Children’s Mercy Kansas City (CM) campuses, allowing for staff expansion in response to increased use by KUCC members. By centralizing operations and reagents between KUCC consortium members, all services are available to KUCC investigators on all campuses at a greatly reduced time and cost. The Cancer Center support of the TGTSR allows for the development of specific initiatives relevant to cancer research. For example, certain transgenic methods and mutations are particularly relevant to cancer studies, including tissue specific transgene expression and subtle mutations that recapitulate clinically identified variants and somatic mutations. Strategic investments by the KUCC have allowed for acquisition of new instrumentation and support to develop novel CRISPR mutagenesis methods in the zygote and in cell lines. The integration of these continually evolving methods into the ‘toolbox’ of the TGTSR greatly accelerates the development of animal and cell models of cancer, while also reducing costs to KUCC researchers on all campuses.