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), intransgenic and knockout mutant mice, and NAD(P)H oxidase mice (gp91phox and p47phox knockout), as wellas oxygen radical-related biochemistry and molecular biology. In addition, Dr. Chan will provide his expertisein primary cell culture of neurons (Project 2), astrocytes (Project 3), and endothelial cells (Project 1) fromrodents (transgenic, knockout, and genetically-matched wild-types). This Core will serve as a centralizedfacility for set-up, for characterization, and for quality control of these primary cell cultures from wild-typeanimals, as well as SOD1 and SOD2 transgenic and knockout mutant mice. In addition, HSP70 transgenicand PARP-1 knockout mice will be bred and genotyped for Project 3 and Project 2, respectively This Corewill 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 biochemicalassays in mice and in primary cell cultures. Finally, this Core will generate and characterize the doubletransgenic/knockout mice (SOD1+/- / gp91phox-/-, SOD2+/- / gp91phox-/-, HSP70+/+ / SOD2-/+) for Projects 1and 3. Thus, the Transgenic Animal Core provides a unique vehicle for the projects to achieve the researchgoals employing various transgenic and knockout animals.
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