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

职业：基于 DNA 分子弹性的 DNA 分离和突变筛选

• 批准号: 0747164
• 负责人: Di Gao
• 金额: $ 40万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0747164&HistoricalAwards=false
• 关键词: CAREER; DNA; Separation; Mutation; Screening

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.

[0747164]这项由化学和生物分离项目颁发的职业奖支持匹兹堡大学高迪教授开发创新的DNA分离和突变筛选技术，这些技术与基于电泳的技术根本不同。DNA分离和突变检测将基于在电场的影响下拉伸DNA链并将其从固体表面拉出。本研究将利用双偏振干涉测量技术研究DNA分子在电动拉伸时的熵弹性。重点将放在DNA在低力（小于0.1 pN）状态下的弹性特性，单链DNA （ssDNA）由自身折叠形成的二级结构，以及碱基对不匹配的双链DNA （dsDNA）。从这些研究中获得的DNA分子的力延伸关系将用于设计和开发基于其粘弹性特性的创新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目前DNA分离技术在几个应用中的局限性，包括(i)染色体大小的DNA分子的分离，（ii）在大DNA片段中筛选基因突变，以及（iii）长读取长度的DNA测序片段的分离。这项研究将展示一种基于粘弹性特性分离生物聚合物的新模式，这也可能应用于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征量身定制个性化医疗的承诺。除了培养研究生和本科生外，本提案的教育部分包括三个部分：(i)为化学工程研究生和大四本科生开发分离过程中的生物聚合物物理课程；（ii）通过与中国清华大学合作，为本科教育开发一个国际实地研究模块，重点关注基因研究的伦理和社会问题的国际观点；（iii）通过与匹兹堡的鲍德温高中和威斯汀豪斯高中合作，开发高中课程单元和研讨会，向高中和未被充分代表的大学前少数民族学生展示科学和创新，其中几乎100%的非洲裔学生。