The Dartmouth Institute for Biomolecular Targeting (bioMT) infuses mechanistic investigations with a sophisticated awareness of disease pathology and therapeutic need, enhancing the quality of even the most fundamental research. At the same time, it helps orient mechanistic investigations towards long-term translational goals for complex diseases such as cancer and infections. In phase I, our progress was strong. All six of our research project leaders (RPL) with more than two years’ support have received R01-equivalent funding. Our cores provide unique protein biochemistry resources and ‘navigators’ to access microscopes campus-wide. Our cores and seminars have created a vibrant and interdisciplinary community. Here, we propose to deploy phase II COBRE and institutional program enrichment funds to build on this foundation and fill key gaps to prepare bioMT for the transition to sustainable COBRE independence. Aim 1 is to increase our cohort of funded bioMT investigators to fill strategic roles in our research landscape. Our two most recent RPLs (3–15 months’ support) are continuing into phase II, joined by two outstanding new hires. Their projects explore basic signaling and immunological mechanisms with potential relevance to therapeutic targets ranging from respiratory infections to cancer progression and metastasis, interconnected by shared scientific and technical interests. All receive guidance from dedicated mentoring dyads to assist in career advancement and independent extramural funding, and all receive support from responsive scientific cores offering state-of-the- art technologies directly relevant to their bioMT research projects. With institutional support, we will also hire five new faculty members, aligned with our theme of discovering and exploiting molecular targets, and selected to enhance thematic subgroups. As starting RPLs graduate, we will recruit new hires and other potential candidates for EAC consideration as replacements. Aim 2 is to enhance our core facilities and prepare them for a transition to COBRE independence during phase III. The cores are fully staffed and have invested heavily in phase I instrumentation. We will partner to add key new technologies based on user input, including parallel protein expression, mass spectrometry, advanced microscopy, and cryoEM. Cost-recovery models will be developed for incremental deployment in phase III to enable core financial independence. Aim 3 continues enriching our community, including mini-symposia and pilot awards to foster new multi-PI and program-project applications, which will contribute to core sustainability. Overall, these aims will leverage proven COBRE strategies – junior faculty hiring, extramural and academic mentoring, excellent administrative and scientific core support, and interdisciplinary community building – to enhance bioMT’s scientific impact in targeted areas of need. This will prepare us for the transition to phase III funding and ultimate independe...