CAREER: Real-Time Transport Phenomena of Non-Viral DNA Nanocarriers in Live Cells: A Research and Education Program

Justin S.HanesBES-0346716这项拟议的研究试图通过使用成像技术和粒子跟踪软件来量化粒子通过潜在的细胞内速率限制屏障的速率和效率。其具体目的是：(1)研究基因载体的物理化学性质对多种重要的非病毒基因载体在人的原代肌肉和内皮细胞中时空依赖性的细胞内转运和积累的影响；(2)确定导致基因载体细胞内转运现象的生物学和/或生物物理机制；(3)确定将重要的生物活性配体组合附着在基因载体表面对载体细胞内转运的影响，并将其与基因表达相关联。从拟议的研究中获得的潜在的科学见解可能为改进设计新的基因载体提供合理的基础，新的基因载体能够更有效地输送到细胞核。在单个载体的基础上确定不同载体群体的运输行为的能力是至关重要的，因为应用于细胞的基因载体中只有很小一部分最终到达细胞核。此外，还提出了两项创新的教育举措，旨在招募和留住年轻人，特别是贫困和代表性不足的学生，进入工程学职业。第一个是为本科生开设的课程和研究相结合的课程，重点是技术交流技能和独立研究。第二个是一个高中外展研究项目，目标是来自贫困家庭的学生，或者是理工科代表不足的群体的学生。

Justin S. HanesBES-0346716This proposed research seeks to quantify the rates and efficiencies of particle transport through potential intracellular rate-limiting barriers by using a imaging technique and particle tracking software. The specific aims are: (1) to investigate the effects of gene carrier physicochemical properties on the spatially- and temporally-dependent intracellular transport and accumulation of a variety of important non-viral gene carriers in human primary muscle and endothelial cells, (2) to determine the biological and/or biophysical mechanisms responsible for the intracellular gene carrier transport phenomena, and (3) to identify the effects of attaching combinations of important bioactive ligands to the gene carrier surface on the intracellular transport of the carriers, and correlate this with gene expression. The potential scientific insights to be gained from the proposed research may provide a rational basis for the improved design of new gene carriers capable of more efficient delivery to the cell nucleus. The ability to determine the transport behavior of a heterogeneous population of carriers on an individual carrier basis is critical since only a minute fraction of gene carriers applied to a cell ultimately reaches the nucleus. In addition, two innovative educational initiatives designed to recruit and retain young people, especially underprivileged and underrepresented students, into engineering careers are proposed. The first is a combined coursework and research program for undergraduates with an emphasis on technical communication skills and independent research. The second is a high school outreach research program targeted toward students that come from underprivileged families, or that are part of underrepresented groups in science and engineering.