CAREER: DNA Separation and Mutation Screening Based on the Elasticity of DNA Molecules

0747164 Gao化学和生物分离项目的CAREER奖支持匹兹堡大学高迪教授的工作，以开发创新的DNA分离和突变筛选技术，这些技术与基于DNA的技术有着根本的不同。DNA的分离和突变检测将基于在电场的影响下拉伸DNA链时将其从固体表面拉下来。 本研究将利用双偏振干涉技术研究DNA分子在电动拉伸下的熵弹性。 重点将放在低力（小于0.1 pN）制度，单链DNA（ssDNA）的二级结构，通过折叠形成的DNA的弹性特性，和双链DNA（dsDNA）与碱基对错配。从这些调查中获得的DNA分子的力-延伸关系将用于设计和开发基于其粘弹性的创新DNA分离和突变筛选过程。从这一发展演变而来的过程将被应用于突破DNA分离技术在几个应用中的当前限制，包括（i）染色体大小的DNA分子的分离，（ii）大DNA片段中基因突变的筛选，以及（iii）具有长读取长度的DNA测序片段的分离。该研究将展示一种基于粘弹性分离生物聚合物的新范式，该范式也可应用于RNA和蛋白质。该项目的成果可能具有实际意义，使遗传分析成为标准体检的一部分，并实现根据个人独特的遗传特性提供个性化医疗服务的承诺。除了培训研究生和本科生外，本建议的教育部分包括三个部分：（i）为化学工程研究生和高年级本科生开设分离过程中的生物聚合物物理学课程，（二）通过与清华大学合作，为本科教育开发国际实地研究模块，重点关注遗传研究伦理和社会问题的国际观点通过与鲍德温高中（Baldwin High School）和匹兹堡西屋高中（Westinghouse High School）合作开发高中课程单元和举办讲习班，（iii）向高中推广科学和创新，并让未被充分代表的少数民族大学预科学生接触科学和创新，匹兹堡的学生几乎100%为非洲裔美国人。

0747164GaoThis CAREER award by the Chemical and Biological Separations program supports work by Professor Di Gao at University of Pittsburgh to develop innovative DNA separation and mutation screening technologies that are fundamentally different from electrophoresis-based ones. Separation of DNA and mutation detection will be based on pulling DNA strands off a solid surface upon stretching them under the influence of an electric field. The research will investigate the entropic elasticity of DNA molecules upon electrokinetic streching using a dual polarization interferometry technique. Focus will be placed on the elastic properties of DNA in low force (less than 0.1 pN) regimes, single-stranded DNA (ssDNA) secondary structures formed by folding upon itself, and double-stranded DNA (dsDNA) with base-pair mismatches. The force-extension relationship of DNA molecules obtained from these investigations will be employed to design and develop innovative DNA separation and mutation screening processes based on their viscoelastic properties. Processes that evolve from this development will be applied to break through the current limitations of DNA separation technologies in several applications including (i) separation of chromosome-sized DNA molecules, (ii) screening of gene mutations in large DNA fragments, and (iii) separation of DNA sequencing fragments with long read lengths. The research will demonstrate a novel paradigm of separating biopolymers based on their viscoelastic properties, which may also be applied to RNA and proteins. The outcome of the project may have practical significance in enabling genetic analysis to become part of a standard physical examination and to realize the promise of personalized medical care tailored to an individual's unique genetic identity. In addition to training graduate and undergraduate students, the education component of this proposal consists of three parts: (i) development of a course on biopolymer physics in separation processes for chemical engineering graduate and senior undergraduate students, (ii) development of an international field study module for undergraduate education focusing on international views of ethical and social issues of genetic research through collaboration with Tsinghua University, China, and (iii) outreach to high school and exposure of underrepresented minority pre-college students to science and innovations through development of high-school course unit and workshop by collaboration with Baldwin High School and Westinghouse High School in Pittsburgh with a nearly 100% African-American student population.