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

这项拟议的研究旨在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限速屏障的速率和效率。具体目标是：(1)研究基因载体的理化性质对人体原代肌肉和内皮细胞中多种重要非病毒基因载体的时空依赖性细胞内转运和积累的影响；(2)确定细胞内基因载体转运现象的生物学和/或生物物理机制；(3)确定重要生物活性配体与基因载体表面的结合对载体胞内转运的影响，并将其与基因表达联系起来。从所提出的研究中获得的潜在科学见解可能为改进设计能够更有效地递送到细胞核的新基因载体提供合理的基础。在单个载体的基础上确定异质载体群体的转运行为的能力是至关重要的，因为只有一小部分基因载体应用于细胞最终到达细胞核。此外，还提出了两项创新教育倡议，旨在招募和留住年轻人，特别是贫困和代表性不足的学生，从事工程职业。第一个是针对本科生的综合课程和研究项目，重点是技术沟通技巧和独立研究。第二个是针对来自贫困家庭的学生，或者是科学和工程领域代表性不足群体的学生的高中外展研究项目。

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.