Biomimetic cytoskeleton and advanced microscopy to reveal intracellular DNA dynamics and distributions

项目总结/文摘

Project Summary/Abstract The proposed diversity supplement is to support undergraduate researcher, Juexin Marfai, a Pacific Islander non-native English speaker, and Navy veteran who transferred to University of San Diego from community college during the pandemic; and, in turn, to enhance the diversity of the health-related workforce. The proposed supplement to support undergraduate researcher Juexin Marfai will contribute to and advance 2 Aims of our parent R15: (Aim 1) Design in vitro active cytoskeleton networks that exhibit tunable activity for probing non- equilibrium dynamics and rheological properties; and (Aim 2) Determine transport and conformational dynamics of linear and circular DNA within active cytoskeleton networks. The supplement will build on Aim 2, beyond the initial scope of the grant by incorporating in situ enzymatically-driven alterations to the DNA topology. To achieve these goals, we will pursue two Aims: (Aim S1) Perform macrorheology measurements on networks designed in Aim 1 to complement measurements using active and passive microrheology; and (Aim S2) Perform Aim 2 experiments to determine transport and conformational dynamics of DNA molecules with different topologies and which transition their topologies from circular to linear. As an undergraduate researcher, Marfai has developed expertise in preparing and characterizing DNA molecules, quantifying the rheological properties of composites comprised of DNA, and studying how the activity of DNA-cutting enzymes causes time-dependent rheological properties of DNA composites. As a senior undergraduate, Marfai will continue to hone these technical skills while applying them to build upon the Aims of our parent grant. Further, we will focus on training Marfai in a broad range of operational and professional skills which will prepare her to accomplish her goal of earning a MD/PhD to continue in biomedical research. We will work with Juexin Marfai to help her achieve her short-term and long-term goals. Short-term goals include: building her health-related research skillset; broadening her professional network and identifying new mentors; and participating in informational interviews with MD/PhD biomedical researchers. Long-term goals include: publishing 2 more papers on her undergraduate research; securing a postbaccalaureate research position in the PI’s lab or in the lab of one of our collaborators; writing a competitive predoctoral fellowship application (either an F30 or F31 NIH fellowship).