The purpose of this Core is to make available various transgenic and knockout mutants for Projects 1, 2, and 3 of the program. One particular strength of this Transgenic Animal Core is that Dr. Chan (Core Leader) will provide his expertise to the other projects in the use of superoxide dismutases (SOD1, SOD2), in transgenic and knockout mutant mice, and NAD(P)H oxidase mice (gp91phox and p47phox knockout), as well as oxygen radical-related biochemistry and molecular biology. In addition, Dr. Chan will provide his expertise in primary cell culture of neurons (Project 2), astrocytes (Project 3), and endothelial cells (Project 1) from rodents (transgenic, knockout, and genetically-matched wild-types). This Core will serve as a centralized facility for set-up, for characterization, and for quality control of these primary cell cultures from wild-type animals, as well as SOD1 and SOD2 transgenic and knockout mutant mice. In addition, HSP70 transgenic and PARP-1 knockout mice will be bred and genotyped for Project 3 and Project 2, respectively This Core will also provide expertise in the molecular analysis of gene expression (PCR, RT-PCR, real-time RT-PCR), gel electrophoresis, and enzymatic activity of CuZnSOD, MnSOD, NAD(P)H oxidase, and biochemical assays in mice and in primary cell cultures. Finally, this Core will generate and characterize the double transgenic/knockout mice (SOD1+/- / gp91phox-/-, SOD2+/- / gp91phox-/-, HSP70+/+ / SOD2-/+) for Projects 1 and 3. Thus, the Transgenic Animal Core provides a unique vehicle for the projects to achieve the research goals employing various transgenic and knockout animals.
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