Grant: Research Tools and Instruments

<p>Light scattering (LS) enables analysis of dynamic changes in the absolute mass, size and shape of objects from 0.3 nm (~1 kDa) to 10 &mu;m (1 MDa) including extracellular vesicles, proteins and multimolecular complexes. This technology can be used either with solutions in a microcuvette or in-line with any column to investigate eluting complexes. Together this allows highly accurate analysis of the interactions between these objects and their sensitivity to changes in their molecular environment (pH, co-factors, chemical). This nanoscale resolution of dynamic changes in objects enables a remarkable variety of transformative applications.<br /> Three labs at the University of Ottawa have an urgent need for LS. Dr. Gibbings studies extracellular vesicles called exosomes to understand the functions of this surprising system of intercellular communication as well as its biotech applications in drug delivery and diagnostics (e.g. <em>Nat.Cell Biol</em> 2009, <em>Nat.Struct.Mol.Biol.</em> 2012, <em>RNA</em> 2014, <em>EMBO Reports</em>, in revision). LS is critical to provide robust measures of exosome quantity, aggregation and purity. This is essential to confidently attribute biological effects to particular types of nano-sized extracellular vesicles. At present the projects of 6 HQP depend upon a very old light scattering instrument for which parts are unavailable when it inevitably breaks.</p> <p >The Figeys and Couture labs dissect the workings of molecular machines in vitro (e.g. <em>Genes and Dev</em>, <em>Cell Reports</em> 2015, <em>Science</em>, <em>PLoS Genetics</em> 2014, <em>PNAS</em> 2013). First, LS would allow them to accelerate 20-100-fold the screening of conditions for purification or crystallization of proteins, a major bottleneck in their research. Second, after protein purification, LS would allow them to analyze the assembly and interactions of molecular machines composed of proteins, nucleic acids or lipids as well as their sensitivity to co-factors and environment. The current obsolete LS instrument cannot perform many of these analyses (absolute mass, shape, poor sensitivity). The proposed LS would greatly accelerate research training of approximately 16 HQP and provide a transformative new suite of tools to their study of molecular machines. These groups also lack an optimal tool to extract high quality proteins. To address this challenge to analyzing molecular machines with LS and other technologies, we are requesting a freezer mill that allows mechanical disruption in liquid nitrogen. <br /> <br /> These instruments will be housed and maintained in the Common Equipment Core that provides maintenance and funds service contracts. This will maximize instrument lifespan. In addition, four dedicated Core staff publicize transformative new equipment and train HQP to use it. This will maximize exposure of HQP beyond the present applicants to cutting-edge technology widely used in industry thereby promoting future innovation and employment in the knowledge economy.</p>