SCRIPPS RESEARCH INSTITUTE, THE
Grant: $379,800 - National Institutes of Health - Sep. 8, 2009
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Award Description: The overall purpose of the award is to conduct a lentiviral shRNA screen in human T cells and identify novel genes that are required for T cell receptor signaling and T cell activation. In year 1, we will conduct the full-scale 15,000 shRNA screen. We have already established all required technology and completed an on-line proof-of-concept screen of 894 shRNAs. Following its completion, we will identify, pick and retest the hits off-line to identify confirmed, reproducible hits for further study. In year 2, we will follow our hit prioritization flowchart (Fig. 2 of the original proposal) to prioritize confirmed hits by novelty and confidence. For hits where only 1 shRNA confirmed, we will generate and test a total of 6 independent shRNAs off-line to maximize the probability that at least 2 shRNAs per each hit gene achieve sufficient knockdown. This will unveil those hit genes where at least 2 independent, validated shRNAs confirm. Next, we will confirm knockdown via qPCR/IB and then use our available in-house and public data to prioritize the hits as described in Fig. 2, generate shRNA-resistant cDNAs for top priority hits and conduct the cDNA rescue studies to select 10 genes where the RNA knockdown phenotype is rescued by expression of shRNA-resistant cDNAs for detailed mechanistic study. If time permits, we will begin to analyze how modulation of these 10 genes affects TCR signaling in Jurkat cells.
Project Description: As defined in Award Description field.
Jobs Summary: We received the award within the past month. It allowed the PI to retain one postdoctoral fellow who would otherwise have been terminated due to funding problems arising from the economic crisis. In addition, we have started the advertizing process to identify and recruit the to-be-appointed technician and ordered the supplies for year 1 listed in the budget justification. (Total jobs reported: 2)
Project Status: Less Than 50% Completed
This award's data was last updated on Sep. 8, 2009. Help expand these official descriptions using the wiki below.
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