RUI: Phage-Based Components for Nanoscale Assembly

Haverford College, Haverford PA

Investigators

Abstract

ID: MPS/DMR/BMAT(7623) 0804944 PI: Johnson, Karl ORG: Haverford College Title: RUI: Phage-Based Components for Nanoscale Assembly INTELLECTUAL MERIT: This proposal aims to develop the filamentous bacterial virus M13 as a component for the assembly of higher-order structures. To accomplish this the PI proposes the following specific tasks: (1) Systematically explore relationships between fluorochrome density, brightness, and quenching for single-color fluorochromes linked to the phage protein coat in order to optimize the ability to image and track individual phage. (2) Use cross-linking chemistries to link FRET (Forster Resonance Energy Transfer) dye pairs and other functional groups to phage and study energy absorption and transfer along the extended protein-DNA backbone in order to lay the groundwork for applications involving photonic energy absorption and transfer. (3) Develop a collection of phage that bind specifically to diverse substrates such as nickel, plastics, silicon, or semiconductor materials in order to target phage SAMs (self assembled monolayers) to unique surfaces. (4) Develop ways to join phage using biomolecular affinity interactions, such as biotin-streptavidin binding or peptide coiled-coil formation, to link phage to each other and around hubs in defined geometries in order to build on these SAM ?foundations?. (5) Study the structural bases of wool-like and cable-like phage networks using fluorescence, atomic force, and electron microscopies. (6) Extend these aggregate measurements to the measurement of individual phage spectral intensities using digital fluorescence microscopy to validate the use of individual pH-sensitive-dye-labeled phase particles (pHages) as tools for the measurement of pH values on ultrafine spatial scales and incorporate these sensors into our higher order assemblies. BROADER IMPACTS: This RUI project will take place at Haverford College, a small liberal arts institution with a diverse student body and a strong tradition of undergraduate student involvement in research. The liberal arts environment is a rich training ground for the nation?s future leaders in science and medicine, and the opportunities made possible by this project will expose students at all levels to the research process. These effects will be magnified by outreach activities involving secondary school students and their teachers through both off-campus talks and on-campus workshops, and will enhance the research-rich environment at the College through support of this interdisciplinary project. Between 4-6 students per year will be involved in accomplishing the objectives of this proposal. The research team will include both underclass students and seniors in the process of doing science over the summer and during academic terms. The training aspects of this proposal develop the human component of US scientific infrastructure by reaching students at a critical point in their careers where they are learning new things and making decisions about future directions. These students will be full participants in this project by their involvement in these experiments, their presentation of their work at national meetings, and their co-authorship on publications in leading journals describing their contributions to this shared endeavor. In a previous RUI grant to the PI involving a total of 30 students, 70% of the undergraduate trainees were female and 43% were members of underrepresented groups. The current project expects to continue these efforts to broaden participation in the nation's scientific enterprise.