Molecular Resources Core

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Summary: Core C, the Molecular Resources Core, will provide molecular reagents and services to all four projects and will be crucial to the completion of the specific aims of each proposal in the Program Project. The proposal for a Molecular Resources Core is a continuation of the original Core C. The overall function of Core C is to provide molecular reagents and expertise, including generation of deletion and site-directed mutants and incorporation into the desired expression vectors, generation of adenoviral and retroviral constructs for transducfion of primary endothelial cells, genotyping of knockout mice, designing and generating appropriate siRNA based constructs, sequencing and verification of constructs obtained from other investigators, molecular biology training to project personnel, and overseeing and maintaining instrumentation for efficient transfection of endothelial cells. Core C will also provide expertise for proteomic analysis and oxidant measurements. This core will be directed by Dr. Randal A. Skidgel and molecular reagents and services will be provided by a Senior Research Specialist (Dr. Tiffany Sharma), Research Specialist (Debra Salvi) and Research Assistant Professor (Dr. Fulong Tan). Dr. Neil Kelleher, Professor and Director of the Proteomics Center of Excellence, will advise, oversee and carry out proteomic analysis by mass spectrometry. Dr. Marcelo Bonini, Assistant Professor, will provide expertise, advice and will carry out measurement of peroxynitrite and other oxidants to dissect their roles in regulating lung vascular endothelial barrier barrier function. Consolidating these efforts in Core C will result in increased speed and efficiency in accomplishing the research tasks outlined in each of the projects and uniformity of approaches and methods so that results from each project can be compared. In addition, it will allow strict quality control and consistency of molecular reagents to be used by the various projects. Furthermore, an added and important benefit of the centralized Core C will be to markedly decrease expenses compared to a scenario in which each project would generate its own molecular resources. Regular meetings of the Core Leader with the Senior Research Specialist and project leaders will assure coordination and prioritization of the generation of molecular reagents and assure that the desired reagents are provided in a timely manner to the component projects.