Project Summary/Abstract The proposed diversity supplement is to support early career faculty member, Dr. Janet Sheung, a Chinese woman in her 2nd year of a tenure-track faculty position in the W.M. Keck Science Department at Scripps College, a predominantly undergraduate institution similar to University of San Diego; and, in turn, to enhance the diversity of the health-related workforce. The aims of the 2-year supplement are to advance and enhance our R15-funded research, and to arm Dr. Sheung with skills, experience, independence, and a strong network of fellow researchers to establish a robust and innovative biophysics research program at her home institution. Specifically, Dr. Sheung will contribute to and advance the following Aims of our parent R15 grant: (Aim 1) Design in vitro active cytoskeleton networks that exhibit tunable activity for probing non-equilibrium dynamics and rheological properties; (Aim 2) Determine transport and conformational dynamics of linear and circular DNA within active cytoskeleton networks; (Aim 3) Link the time-varying spatial distributions of linear and circular DNA to cytoskeleton network properties and activity. This supplement will also build on Aim 3, beyond the initial scope of the grant, by Dr. Sheung building a novel instrument that combines optical tweezers (OT) and light-sheet microscopy (LSM) to perform simultaneous measurements of cytoskeleton network mechanics and macromolecule (i.e., DNA) dynamics. This instrumentation and method advance will, in turn, place Dr. Sheung’s research lab at the forefront of biomedical and biophysical research, draw in new external collaborators, and set her on a strong trajectory as an independent researcher. To achieve these goals, we will pursue three Aims: (Aim S1) Perform Aim 2 experiments to determine transport and conformational dynamics of linear and circular DNA within active cytoskeleton networks; (Aim S2) Build a novel combined fluorescence light-sheet microscope (LSM) and force-measuring optical tweezers (OT) in Dr. Sheung’s lab to provide unique contributions to our R15 Aims 1-3 and establish a robust undergraduate-driven research program at Dr. Sheung’s home institution; (Aim S3) Perform experiments that contribute to Aims 1-3 of the parent R15 with instrumentation established in Dr. Sheung’s lab in Aim S2. We will build on the strong mentoring and collaboration relationship we have established with Dr. Sheung, to help her achieve her short-term and long-term career goals. Short-term goals include: (i) gaining the expertise to construct a light-sheet microscope and optical tweezers in her own lab; (ii) developing strategies to effectively mentor undergraduate and high school students in summer research projects; and (iii) receiving federal funding for her research. Long-term goals include: (i) establishing a fruitful independent interdisciplinary research program accessible to undergraduates from multiple disciplines, (ii) building a network of external c...