Genetics / -Omics

Gene Targeting & Knockout Shared Resource

Director

Demetri D. Spyropoulos, PhD
Associate Professor of Pathology & Laboratory Medicine
Phone: 843-792-1625
E-Mail: spyropdd@musc.edu

Website:

http://hcc.musc.edu/research/resources/genetargetingknockout.htm

Description:

This Shared Resource consists of 4 facilities: Transgenic, Gene Targeting and Knockout, Xenograft, and Carcinogenesis. Transgenic and Gene Targeting facilities are essential genetic tools for studying mammalian gene functions in vivo, by testing the over-expression and loss-of-function mutations in animal development and in tumor susceptibility, respectively. In addition, Transgenic technology can be used in generating tissue-specific expression vehicles that can be utilized in creating conditional knockouts, either by Cre recombinase or by siRNA expression. Xenograft of human cancer cells or tissues into immune deficient mice is an indispensable step in correlating in vitro cell biology studies of cancer biology with tumorigenicity in vivo. The tumor grown in mice can also serve as a realistic test model for new therapeutics. For studying specific cancer types, the Carcinogenesis facility is able to generate several organ-specific cancers in rodents; they are invaluable tools for testing the function and therapeutic values of specific genes and therapies. The Cancer Animal Model Shared Resource combines the existing expertise of these four facilities to form a comprehensive service to assist cancer researchers at HCC.

The overall goal of Gene Targeting and Knockout component of the Animal Models Shared Resource is to provide the means by which researchers can learn and apply cutting edge technology to the molecular analysis of mammalian gene function. The specific aim of this facility is to create "knockout mice" through the utilization of DNA-, stem cell-, and embryo-manipulation procedures. Molecular cloning techniques are employed to clone and manipulate DNA sequences. Homologous DNA recombination in cultured embryonic stem (ES) cells is employed to generate "targeted" ES cells (i.e., ES cells carrying the replacement of specific chromosomal DNA sequences with cloned DNA sequences). Embryo manipulation techniques involving the targeted ES cells are employed to generate chimeric mice, which are then used to generate the knockout mice.

Access:

Open to MUSC faculty, students and associated technical personnel.