CRISPR, VECTOR AND TRANSGENIC MOUSE CORE

The overall objective of the CRISPR, Vector and Transgenic Mouse Core (CVTMC) is to provide Diabetes
Research Center (DRC) affiliate investigators at the University of Washington (UW) and the Greater Seattle
area with state-of-the-art vectors necessary to edit, monitor, or alter expression of genes and RNAs of interest
in cultured cells, tissues, and animals. This well-used Core has evolved considerably since the last competitive
renewal, based on the changing needs of affiliate investigators. New and more efficient CRISPR methods have
been developed, use of viral vectors containing cell type-selective promoters has been greatly expanded, and
more sophisticated virus production and purification methods have been implemented. These newer services
are highly used, and now provide the bulk of the Core’s work. Conversely, services that are less cutting-edge
and are no longer frequently requested have been removed. The specific aims of the Core are to provide the
following services to affiliate investigators: (1) Production of CRISPR reagents for genome editing and for
various other applications, such as CRISPR-mediated transcriptional activation and repression; (2) Facilitate
generation of mouse models by CRISPR through collaboration with the Transgenic Resources Program at the
UW; (3) Facilitate generation of cell lines by CRISPR in collaboration with the Institute for Stem Cell and
Regenerative Medicine (ISCRM) at the UW; (4) Production of adeno-associated viral (AAV) and lentiviral
vectors for use in animals, tissues and cells; (5) Development of genotyping methods to validate edited mice
and cell lines; and other specialized molecular biology, such as cloning and PCR; and (6) Consultation at each
step and training of affiliates and their trainees in the Core’s methods. The Core has been highly productive in
the current funding period and has added new services that are expected to significantly increase productivity
and usability for DRC affiliate investigators to meet the Center's goal to enhance research in diabetes, obesity
and related disorders in the Greater Seattle area and beyond.

Grant Number: 5P30DK017047-50
NIH Institute/Center: NIH
Principal Investigator: Karin Bornfeldt