Abstract/Project Summary RODENT GENETIC ENGINEERING LABORATORY SHARED RESOURCE (RGEL) The Rodent Genetic Engineering Shared Resource Laboratory (RGEL) is a new PCC Shared Resource that offers innovative, comprehensive rodent genetic engineering services that include: CRISPR/Cas9-mediated production of mutant mice, generation of transgenic mice by pronuclear injection of DNA or BAC-DNA, gene targeting in mouse C57BL/6 embryonic stem (ES) cells, generation of fully ESC-derived mice using tetraploid complementation and chimeras with genetically altered ES cells, isolation and establishment of genetically-modified ES cells from existing strains for serial gene targeting, and re-derivation and cryopreservation services for both embryos and sperm. Under the direction of Dr. Sang Yong Kim, who has extensive experience in mouse developmental biology and all aspects of manipulation of the mouse germ line, RGEL is used extensively by PCC investigators to make novel genetically engineered mouse models of cancer (GEMMs), as well as to rederive and store various mouse strains. RGEL accomplishes its goals by means of four Specific Aims: Aim 1) To develop and apply state-of-the-art technology and expertise in support of rodent genetic engineering; Aim 2) To provide highly efficient, cost-effective services in DNA pronuclear injection, gene editing using CRISPR-Cas9 and related systems, blastocyst injection (ES cell, iPS cell), ES cell gene targeting and expansion, embryo tetraploid complementation, embryo rederivation, embryo & sperm cryopreservation, in vitro fertilization (IVF) and transgenic animal genotyping; Aim 3) To leverage expertise in experiment genetics, cell/molecular research and systems biology for genotypic/phenotypic analyses; Aim 4) To provide scientific leadership for a collaborative center of service, technology and expertise in experimental genetic mutation resources.