Grant: Research Tools and Instruments

Mechanistic studies at the molecular level require the manipulation of gene expression to modify protein abundance (i.e. increase or decrease expression). This manipulation can be done using a variety of methods, but all methods result in the transfer of RNA or DNA directly into cells. Transfection must be efficient with low toxicity to the cells in order to generate high quality, robust and reliable data. The applicants' research programs are highly dependent on transfection technology for their NSERC-funded research programs. These programs focus on determining the pathways that regulate B lymphocyte development and activation, tracking protein localization to better understand receptor-mediated activation of cells in response to trace amines, elucidating the mechanisms behind the generation of foam cells from macrophages, and understanding the mechanisms that regulate PIWI RNA-mediated gene silencing. Support for the system currently available to the applicants is being discontinued by the manufacturer, leaving these groups without an essential tool for advancing their research programs. The loss of this system will completely impede the research progress of these groups. Thus, they are requesting funds to acquire a new and superior transfection system that will allow efficient transfection of both cell lines and primary cells with low toxicity. The ability to transfect primary cells and hard-to-transfect cell lines with low toxicity is a major advantage of the Nucleofector system compared to other electroporation systems. Acquisition of this equipment is critical for the applicants to continue to make progress in their research.