Transgenic and Gene Targeting

The Transgenic and Gene Targeting Shared Resource (TGTSR) is an important shared resource providing centralized technical services for the design, production and analysis of genetically engineered rodents and pluripotent stem cells for researchers at The University of Kansas Cancer Center.

The TGTSR has 5 aims:

1. Provide technical expertise in experimental design for The University of Kansas Cancer Center projects using mice and pluripotent stem cells
2. Provide technical expertise for design, validation and use of genome editing using CRISPR/Cas9 tools
3. Provide technical support for genetic alteration of mice and pluripotent stem cells
4. Provide support for mouse strain development, maintenance and cryopreservation
5. Educate cancer center researchers and trainees on new and emerging technologies for genetically altered animal models and manipulation of pluripotent stem cells

The complex etiology of tumor development and physiological responses to cancer and cancer therapeutics require the use of animal models. As such, genetically altered mouse models are significant tools for the researchers at the cancer center. The production and analysis of such models ultimately lead to better understanding of biological processes of tumorigenesis, as well as in vivo models for diagnostic testing and cancer treatment.

The TGTSR provides comprehensive services to support the cancer center research related to genetic manipulation of the mouse and stem cell genomes. This shared resource uses front-end technologies, including genome editing via CRISPR/Cas9 tools for genetic manipulation of the mouse in vivo. The TGTSR has dedicated laboratory space for molecular biology, cell culture, embryo manipulation and animal housing for comprehensive support for making and using novel genetically modified mice.

Learn more about the Transgenic and Gene Targeting Institutional Facility.

View the Transgenic and Gene Targeting Brochure.

Large templates for CRISPR-mediated homology-directed repair

The TGTSR has developed optimal methods for using large double-stranded donor DNA as templates for CRISPR-mediated homology-directed repair (HDR) in vivo.  This system allows for the targeting of large DNA elements to specific regions of the genome. The mouse genetics field is still determining the best methods for performing such CRISPR-mediated manipulations in vivo.  Recent studies using large single-strand DNA templates have shown that such methods result in unacceptably high rates of small undesired deletions. To these ends, the shared resource team has focused its efforts on optimizing the use of long double-strand DNA templates, derived from plasmids, for targeting relatively large DNA elements (200bp-~8kb). These procedural advancements include the use of optimal size of arms of homology and specific preparation methods of the DNA template for injection. These efforts have resulted in a robust, practical, cost-effective and generalizable means of CRISPR-mediated HDR using large double-stranded DNA templates in vivo.

Cite the Cancer Center Support Grant

This resource is funded by The University of Kansas Cancer Center Support Grant (CCSG) awarded by the National Cancer Institute (P30 CA168524). Publications that have utilized facility resources, services or scientific data generated by the resource should acknowledge the resource and cite the NCI CCSG grant.