Molecular Tools Core

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The goal of the Molecular Tools Core (MTC) is to provide biomolecular tools to the bioMT faculty to allow them to better achieve their research goals. During the first funding period the MTC assisted over 200 researchers within the greater biomedical community and played a central role in the advancement of the bioMT junior research project leaders (RPL) to externally funded independent investigators. Our success is based on the centralization of the equipment and expertise for providing high-quality purified proteins and other biomolecular tools, dedicated laboratory work by our expert staff, as well as training of users in the utilization of state-of-the- art instrumentation maintained within the core. To complement the expertise of our faculty Director, we recruited a highly experienced Lead Scientist to oversee the operation of the MTC and to provide expert protein biochemistry support and consultation. Her experience in protein production has served her well particularly as a consultant, providing advice from sequence analysis to expression systems to purification protocols to both experienced and novice users. We have established efficient pipelines for protein production in E. coli and mammalian cells and optimized purification schemes. In collaboration with the Molecular Interactions and Imaging Core (MIIC), we have developed protocols for biochemical and biophysical characterization and QA/QC of the final products. Additionally, during phase 1, the MTC created a bioMT Service Center for chargeback costs of supplies, media and reagents related to eukaryotic cell protein production starting October 1, 2020. In the next funding period, we will continue to provide valuable resources to all four RPLs as detailed in Aim 1. One vital feature will be the addition of standard methods for protein production using insect cell lines (Sf9, Hi5). Aim 2 is to enhance the efficient operation of the core, streamlining the day-to-day operations and overall throughput to better serve the research community. We will build on our collection of resources, including strains, vectors, in-house produced enzymes and resins. As part of our phase II strategy, we will expand our throughput capabilities with respect with construct design, cloning, expression screenings to rapidly identify protein targets, reducing the burden of the bioMT investigators on protein production. As the number of users of the MTC has expanded, it is imperative that we invest in the efficiency of the operation of the core and maintain a robust user base as we transition to a cost-share center to guarantee sustainability. Another key element of our phase II strategy is sustainability. As outlined in Aim 3, we will develop cost-recovery models. Faculty will not be charged during phase II, but we will share quarterly usage with projected costs, so they can include costs in grant submissions prior to phase III. Looking towards the future, the MTC will also support collaborative pilot projects designed to nucleate multi-project grants. Overall, the phase II plan for MTC will continue to catalyze the success of RPLs and other bioMT faculty, drawing our faculty together and laying the foundation for the Institute’s long- term transition to COBRE independence.