The primary purpose of the Transgenic/Knock-out Mouse Shared Resource is to provide an efficient andeconomical means for producing genetically modified mice for members of the Massey Cancer Center.Genetically modified mice (including transgenic mice, in which customized genes are introduced into thegenome, or 'knock-out/knock-in' mice, in which endogenous genes within the mouse genome arespecifically inactivated or modified) have become an indispensable tool in cancer research. The utility ofsuch models ranges from studies of the basic biology of tumorigenesis and progression to the creation ofgenetically accurate tumor models for the evaluation of novel therapeutic approaches. TheTransgenic/Knock-out Mouse Shared Resource offers the following services:Transgenic mouse production: The core generates at least three founder mice for each transgene.Knock-out/knock-in mouse production: The core can perform all or a subset of the followingprocedures: ES cell electroporation; ES cell culture under appropriate selection conditions; expansion ofdrug-resistant clones; PCR/Southern blot screening of clones for the correct recombination event; reelectroporationwith a ere expression vector to delete floxed selection markers and subsequent rescreeningof clones; injection of targeted ES cell clones into blastocysts for chimeric mouse production.For projects in which the core has performed the ES cell targeting, at least two independent ES cellclones are generally injected into blastocysts, and a minimum of five chimeras are generated.Mouse line rederivation: Lines of pathogen-infected mice are rederived by embryo transfer to generatespecific pathogen-free mice.Sperm cryopreservation: Lines of mice are cryopreserved by collection, freezing, and long-term liquidnitrogen storage of sperm. When needed, re-establishment of the line can be accomplished by thawingsperm and performing in vitro fertilization of eggs isolated from WT females.Tail DNA genotyping: The core isolates genomic DNA from tail biopsies (or other tissue) provided bythe investigator and performs PCR genotyping.Consulting services: The core provides consultation on many aspects of transgenic/knock-out mousetechnology, including experimental design, vector design, budgeting for mouse colony maintenance, andpreparation of applications to the IACUC for the use of laboratory animals in research. In addition, thecore staff provides instruction on overall colony management, including breeding strategies, weaning, earpunching, and tail DNA isolation.
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