Core C is Gene Delivery Scientific Core whose goal is to provide Program Investigators with complete in vitro and in vivo gene delivery solutions to facilitate completion of their specific aims. To accomplish this goal, the Core C will channel its activities into three principal areas: a) providing services; b) developing and acquiring new technologies, and c) building synergy with Projects and other Cores. Service component: The Core C will serve a resource for all genetic manipulation needs of Projects and Cores of this Program. Specifically, the Core C will a) achieve economy of scale and quality assurance through centralized generation of recombinant DNA, viral constructs, and genetically modified microorganisms by highly trained personnel; b) provide technical expertise, consultation and training in PCR, RT-PCR, qPCR, DNA sequencing, gene delivery to prokaryotic and eukaryotic cells, constitutive and regulated gene silencing; c) execute gene targeting and CRISPR/Cas and TALEN-mediated gene editing in cultured cells for the needs of PPG projects; d) provide PPG Projects with enabling technologies for the production of genetically modified laboratory animals, such as Bacterial Artificial Chromosome (BAC) modification, generation CRISPR/Cas constructs for genome editing and genotyping of genetically altered animals; e) provide PPG investigators with recombinant protein expression and purification expertise; f) Core C will provide training, experience and expertise in the field of mitochondrial biology, including generation of cells depleted of mitochondrial DNA and assays of bioenergetic status using Seahorse XF-24 Extracellular Flux Analyzer. Academic component: Throughout previous cycles of funding Gene Delivery Core responded to changing needs of the Projects by either acquiring new technologies or developing its own. This is exemplified by a spectrum of custom retro- and -lentiviral vectors developed by the Core C, development of a system for doxycycline-regulated gene expression, development of a system for regulated gene knockdown, by the development of the gene expression system with a two-tier regulation, etc. In the course of the proposed competitive renewal, Core C will fully implement, validate, and make available to a broad scientific community a system for endothelial segment-restricted gene expression. In addition, we plan to establish a system for point mutations knocking-in in endothelial cells. Synergy with Projects and Scientific Cores: The Core C is a convergence point for all Projects and Cores regarding genetic manipulation of pro- and eukaryotic organisms. The Core C also builds synergy with other Scientific Cores through, e.g. procurement of endothelial cell lines from Core B, modifying these cell lines for the expression of fluorescent reporters/sensors for the needs of Core D, and supplying these modified cell lines to Core D for the use and to Core B for distribution to scientific community.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
2P01HL066299-16A1
Application #
9416707
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Program Officer
Xiao, Lei
Project Start
Project End
Budget Start
2017-09-01
Budget End
2018-08-31
Support Year
16
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of South Alabama
Department
Type
DUNS #
172750234
City
Mobile
State
AL
Country
United States
Zip Code
36688
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