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

项目摘要/摘要 拟议的多样性补充是为了支持本科生研究员、太平洋岛民觉欣·马费 非英语母语者，海军退伍军人，从社区转到圣地亚哥大学 在大流行期间，我们将继续在大学里工作；反过来，加强与卫生相关的劳动力的多样性。建议数 支持本科生研究员觉欣·马费的补充将有助于和推进我们的两个目标 亲本R15：(目标1)体外设计活性细胞骨架网络，显示可调节的探查非 平衡动力学和流变性；以及(目标2)确定运输和构象动力学 活跃的细胞骨架网络中的线状和环状DNA。增刊将建立在目标2的基础上，超越 通过将原位酶驱动的改变纳入DNA拓扑结构，初步确定了赠款的范围。要实现 为了实现这些目标，我们将实现两个目标：(目标S1)在设计于 目标1利用主动和被动微观流变学补充测量；(目标S2)执行目标2 测定不同拓扑结构DNA分子输运和构象动力学的实验 并将它们的拓扑结构从圆形过渡到线性。 作为一名本科生研究人员，Marfai在DNA的制备和表征方面积累了专业知识 分子，量化由DNA组成的复合材料的流变性，并研究 DNA切割酶的变化导致了DNA复合材料的随时间变化的流变性。作为一名大四学生 作为本科生，Marfai将继续磨练这些技术技能，同时应用它们来实现 我们的父母拨款。此外，我们将重点培训Marfai广泛的业务和专业技能 这将为她实现获得医学博士学位以继续从事生物医学研究的目标做好准备。 我们将与觉心玛菲一起努力，帮助她实现短期和长期目标。短期目标包括： 培养她的健康相关研究技能；拓宽她的专业关系网，寻找新的导师； 并参加对医学博士生物医学研究人员的信息性采访。长期目标包括： 又发表了两篇关于她本科生研究的论文；获得了一个学士学位后的研究职位 PI的实验室或我们的一个合作者的实验室；编写一份竞争激烈的博士前奖学金申请(或者 F30或F31 NIH奖学金)。

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).