CAREER: Synthetic Design and Biological Study of Novel Polymeric DNA Delivery Vehicles

辛辛那提大学的Theresa Reineke教授将在化学系的有机和高分子化学计划以及分子和细胞生物科学系的生物分子系统计划的支持下，研究羟基的存在、胺中心的数量、阳离子电荷中心的密度、电荷中心的类型和聚合物材料的几何形状如何促进有效的核酸结合，并将确定上述化学和结构特性如何影响活细胞内的细胞毒性、细胞内递送和基因表达。高效无毒的核酸载体的开发已成为基因功能、细胞信号、基因治疗、基因疫苗和植物生物技术等研究领域面临的最重要和最基本的问题之一。由于与基于病毒的传递系统相关的危险，聚合物载体现在显示出巨大的希望。然而，毒性、低传递效率以及缺乏对聚合物结构-生物特性关系的了解等问题阻碍了它们的使用。该提案的目标将通过合成五个不同于上述功能的新型聚合物家族来实现，并通过等温滴定量热法、动态光散射和Zeta电位测量来研究聚合物与DNA的相互作用。Lowry蛋白、β-半乳糖苷酶和荧光素酶基因的表达分析将在各种转染新聚合物的细胞系中进行。分子和细胞生物科学系的有机和大分子化学项目以及生物分子系统项目支持辛辛那提大学的Theresa Reineke教授，她将建立和促进互动指导、合作学习和研究项目，成功地将教学和学术结合起来。这将通过制定一项名为新兴民族科学家(EES)的结构化指导计划来完成，该计划将互动和合作学习技术纳入一门新生化学课程，并在科学研究过程中指导高中生和本科生。这项工作的长期目标和更广泛的影响是制定一种设计模型，可供聚合基因传递领域的研究人员用来为特定细胞类型和应用开发高效和无毒的载体，并向学生传授进行跨学科研究的兴奋，目的是增加科学中代表性不足的群体的留存。参与这一跨学科项目的学生将在从合成化学和聚合物科学到分子生物学的多个领域获得独特的广泛和深入的经验。

With the support of the Organic and Macromolecular Chemistry Program in the Chemistry Division and the Biomolecular Systems Program in the Molecular and Cellular Biosciences Division, Professor Theresa Reineke of the University of Cincinnati will identify how the presence of hydroxyl groups, the number of amine centers, the density of the cationic charge centers, the charge center type, and the geometry of polymeric materials promote effective nucleic acid binding, and she will determine how the chemical and structural properties detailed above affect cytotoxicity, intracellular delivery, and gene expression within living cells. The development of efficient and nontoxic nucleic acid delivery vehicles has become one of the most significant and fundamental problems facing research fields involving gene function, cellular signaling, gene therapy, genetic vaccines, and plant biotechnology. Due to the dangers associated with virus-based delivery systems, polymeric vectors now show great promise. However, problems with toxicity, low delivery efficiency, and a lack of insight into the polymer structure-biological property relationships have hampered their use. The goals of the proposal will be accomplished by synthesizing five novel polymer families that vary in the functionalities mentioned above and by studying the polymer-DNA interactions via isothermal titration calorimetry, dynamic light scattering, and zeta potential measurements. Lowry protein, beta-galactosidase, and luciferase gene expression assays will be performed with a variety of cell lines transfected with the new polymers. The Organic and Macromolecular Chemistry Program and the Biomolecular Systems Program in the Molecular and Cellular Biosciences Division supports Professor Theresa Reineke of the University of Cincinnati who will establish and promote interactive mentoring, cooperative learning, and research programs that successfully unite teaching and scholarship. This will be completed through developing a structured mentoring program called Emerging Ethnic Scientists (EES) that incorporates interactive and cooperative learning techniques into a Freshman Chemistry course and mentoring high school and undergraduate students in the process of scientific research. The long-range goals and broader impacts of this work are to formulate a design model that can be utilized by researchers in the field of polymeric gene delivery to develop efficient and nontoxic vectors for specific cell types and applications and impart to students the excitement of performing interdisciplinary research with the goal of increasing the retention of underrepresented groups in the sciences. Students involved in this interdisciplinary program will gain a uniquely broad and in-depth experience in several areas ranging from synthetic chemistry and polymer science to molecular biology.