Functional DNA Amplicons

我的研究计划的长期目标是进一步开发生物分子固定化方法库中的其他工具。首先，我们的目标是研究滚动圆扩增功能核酸(FNAs)形成的DNA和RNA超结构的性质，包括DNA和RNA适配子和DNA酶，以扩大此类组件作为生物分子固定平台的用途，并将这些组件应用于一系列生物分析设备，在这些设备中，我们将开发它们在各种生物传感、催化和亲和分离应用中的潜力。在新的资助期，我们将追求以下四个短期目标，以巩固我们正在进行的DG资助计划的势头。目标1制造FNA扩增片段：该项目的主要目标是扩大FNA扩增片段的谱系，并评估FNA扩增片段在NFS中的性质。这项工作是该提案的基础，将为3D扩增子超结构的多功能性以及如何控制这些新结构的行为提供重要的见解。AIM 2蛋白质扩增结合物：该项目的目标是通过杂交或适体-蛋白质相互作用将DNA寡聚体修饰的蛋白质结合到扩增子上，并将这些物种用于上述应用(柱子、纸传感器)，从而将扩增子平台扩展到功能DNA物种之外。这将在我们的团队中开辟一个全新的研究领域，并可能在使用固定化蛋白质进行生物分析方面取得一些突破。Aim 3DNA扩增亲和色谱柱：这项工作将解决使用基于溶胶-凝胶的生物亲和色谱柱的一个根本问题，即限制小分子分析。这项拟议的研究将建立在我们最近关于扩增子掺杂柱的报告的基础上，并将扩大用于形成扩增子柱的功能核酸的类型和这些柱的整体应用。目的4利用扩增子的纸基传感器：将生物分子固定在不同的表面上，如纸的纤维素网络，仍然是一个挑战。我们将研究FNA扩增器作为生产多路纸张传感器的平台的使用，并将研究印刷扩增器的特性以及它们在纸表面的行为。意义和影响：拟议的研究计划将开发一种固定生物制剂(DNA适配子、DNAzyme和蛋白质)的新方法，该方法与亲和柱、生物传感器或其他生物活性表面的制作兼容。其结果将是一个高度通用的生物接口平台，它是高度可调的，可以适应多种应用。我们DG资助项目的新发现将创造新技术，并为参与生物活性材料开发的科学家和工程师增加新工具。

The long-term objectives of my research program are to further develop additional tools in the repertoire of methods for biomolecule immobilization. Primarily, we aim to investigate the nature of DNA and RNA superstructures formed from rolling circle amplification of functional nucleic acids (FNAs), including DNA and RNA aptamers and DNA enzymes, to expand the use of such assemblies as a platform for biomolecule immobilization, and to apply these assemblies to a range of bioanalytical devices where we will exploit their potential for various biosensing, catalysis and affinity separation applications. For the new funding period, we will pursue the following four short term aims that build upon the momentum of our ongoing DG funded program.Aim 1 Fabrication of FNA Amplicons: The key goal of this project is to expand the repertoire of FNA amplicon NFs, and evaluate the properties of FNAs within NFs. This work is foundational for the proposal, and will provide important insights into the versatility of 3D amplicon superstructures, and how to control the behavior of these novel structures.Aim 2 Protein-Amplicon Conjugates: This project will aim to extend the amplicon platform beyond functional DNA species by binding DNA-oligomer-modified proteins to amplicons via hybridization or aptamer-protein interactions, and using these species for the applications noted above (columns, paper sensors). This will open up a completely new area of study in our group, and is likely to produce a number of breakthroughs in bioanalysis using immobilized proteins.Aim 3 DNA Amplicon Affinity Columns: This work will address a fundamental problem in the use of sol-gel based bioaffinity columns the restriction to small molecule analytes. The proposed study will build on our recent report on amplicon-doped columns and will expand the types of functional nucleic acids used to form amplicon columns and the overall applications of these columns. Aim 4 Paper-based Sensors Utilizing Amplicons: Immobilization of biomolecules onto heterogeneous surfaces, such as the cellulose network of paper, remains a challenge. We will investigate the use of FNA amplicons as a platform for producing multiplexed paper sensors, and will investigate the properties of printed amplicons and how they behave on paper surfaces. Signficance and Impact: The proposed research program will develop a new method for immobilization of biological agents (DNA aptamers, DNAzymes and proteins) that is compatible with fabrication of affinity columns, biosensors or other bioactive surfaces. The outcome will be a highly versatile, generic biointerface platform, which is highly tunable and can be adapted to multiple applications. The new discoveries from our DG funded program will create new technologies and add new tools to the arsenal of scientists and engineers involved in development of bioactive materials.