The High-throughput Biology Core will express recombinant proteins and protein complexes, purify recombinant proteins to homogeneity, and produce high-titer stocks of lentiviruses for delivery of cDNAs, shRNA and CRISPR/dCAS9 to (epi)genetically manipulate cells in vitro. Furthermore, the Core will support high-throughput screening to identify genes and small molecule inhibitors, respectively, of key signaling pathways that regulate protein functions or phenotypes associated with cellular senescence and/or age-related pathologies in tissues, including the pro-aging senescence-associated secretory phenotype (SASP). The centralization of these services under the direction of an experienced core leader who has been a long standing member of this PO1, highly trained technical staff, and the unique laboratory infrastructure, including robotic pipetting workstations, a multi-label plate readers, a high- content screening reader, small molecule and genetic libraries, minimizes redundancy; maximizes biosafety associated with recombinant DNA technologies; ensures preparation of high-quality biological materials for seamless integration of reagents across projects; preserves the time of project staff to focus on experimentation to test key hypotheses; and enables investigators to use unbiased approaches to identify new genes and small molecules that regulate protein function or cellular processes. Collectively, these attributes of the proposed core will enable high-throughput, economic, and rapidly scalable services to produce high-quality biological materials. Based on progress from the current funding period and usage of the proposed research activities, the core will continue to be an integral component of the overall program by generating many recombinant protein reagents, viral vectors, and small molecule inhibitors that will facilitate long-term programmatic interactions and enable central hypotheses of this program project to be tested.
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