EAGER: Rapid DNA detection with a fluctuating surface-active peptide

0923610U智能优点：在这项建议中，PI解决了对肽折叠动力学可以应用于在选择性结合过程中加速动力学这一概念的早期探索。这一点很重要，因为将特定疾病与特定生物标记(如DNA)相结合的能力正在以比在复杂环境中快速检测这些标记的能力更快的速度增长。在低分子量DNA标记的序列特异性检测方面已经投入了大量的跨学科努力，但这些努力仍然比不上天然生物分子利用动力学来检测高分子量系统中DNA标记的能力。这一探索性的研究思路集中在如何设计一个新的仿生平台，应用加速动力学来开发快速传感技术。为了做到这一点，人们需要平衡折叠和未折叠的多肽种群的能量格局，这涉及到一系列固有的复杂和危险的实验。该方法依赖于一种算法，该算法结合了二元序列设计和自然发生的DNA结合序列来开发新的多肽。这些未经测试的生物分子与现有的生物分子明显不同，现有的生物分子旨在稳定特定的折叠状态。他们将把他们之前开发的可调表面活性多肽设计与DNA结合基序杂交，并测量结合的选择性、亲和力和结合动力学随波动的二级结构的函数。如果这些早期实验成功，它们将为设计具有快速动力学和高选择性结合的多肽奠定分子基础。这种用于分子设计的新范式随后可以打开大门，探索各种基本问题和技术。将这一想法转化为可行技术的下一步取决于(1)将快速动力学转化为灵敏和坚固的传感器的能力，以及(2)将选择性转化为DNA以外的广泛靶的能力。广泛影响：拟议工作的科学成果将有助于基本理解导致动力学增强的合作折叠和结合过程。此外，拟议的探索性工作还处于早期阶段，但成功的实验证明了这一研究思路将导致一代为动态行为而设计的生物分子，而不是静态的锁和钥匙绑定。这些设计可以转化为基于快速搜索复杂混合物的通用生物技术。这一计划的几个要素可以纳入CCNY的外展工作。这些努力为哈莱姆区数学、科学和工程高中(HSMSE)的学生提供了机会，让他们有机会在纽约城市大学的研究生中参与研究，了解跨学科的科学和工程。这一综合研究和教学计划的深远目标与CCNY为高度多样化的学生群体(36%西班牙裔、30%黑人、23%亚裔和11%白人)提供优质教育的使命相匹配。

0923610TuIntellectual Merit:In this proposal, the PI addresses early stage exploration of the concept that peptide folding dynamics can be applied to accelerate the kinetics in selective binding processes. This is important because the ability to couple a particular illness with a specific biological marker, such as DNA, is increasing faster than the ability to rapidly detect those markers in a complex environment. A significant interdisciplinary effort has been invested in looking at the sequence specific detection of low molecular weight DNA markers, but these efforts still do not match the ability of native biological molecules to use dynamics to detect DNA markers in high molecular weight systems. This exploratory research idea focuses on how one can engineer a new biomimetic platform that applies accelerated kinetics to develop rapid sensing technologies. In order to accomplish this, one needs to balance the energy landscape of folded and unfolded peptide populations, which involves an inherently complex and risky set of experiments. The approach relies on an algorithm that combines binary sequence design and naturally occurring DNA binding sequences to develop novel peptides. These untested biomolecules are distinctly different from existing biomolecules, which are designed to stabilize a particular folded state. They will hybridize their previously developed tunable surface active peptide design with a DNA binding motif and measure the selectivity, avidity and kinetics of binding as a function of the fluctuating secondary structure. If these early stage experiments are successful, they will establish the molecular basis for designing peptides with fast kinetics and highly selective binding. This new paradigm for molecular design could then open doors to explore a variety of fundamental questions and technologies. The next step to transform this idea into a viable technology depends on (1) the ability to translate the fast kinetics into a sensitive and robust sensor and (2) the ability to translate the selectivity to a wide range of targets beyond DNA.Broader Impact:The scientific outcomes of the proposed work will contribute to the basic understanding of cooperative folding and binding processes that result in enhanced kinetics. Moreover, the proposed exploratory work is in its early stages, but successful experiments demonstrating the research idea will lead to a generation of biomolecules designed for dynamic behavior rather than static lock and key binding. These designs can be transformed into a general biotechnology based on quickly searching complex mixtures. Several elements of this program could be integrated into outreach efforts at CCNY. These efforts provide opportunities for students from the High School of Math, Science and Engineering (HSMSE) in Harlem to graduate students in CUNY to participate in research and learn about interdisciplinary science and engineering. The far reaching goals of this integrated research and teaching plan match CCNY's mission to provide quality education to a highly diverse student body (36% Hispanic, 30% black, 23% Asian and 11% white).