Functional DNA Amplicons

功能性 DNA 扩增子

• 批准号: RGPIN-2018-05587
• 负责人: Brennan, John
• 金额: $ 9.32万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748242
• 关键词: Functional; DNA; Amplicons

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.

我的研究计划的长期目标是进一步开发生物分子固定方法中的其他工具。 首先，我们的目标是研究由功能核酸（FNA）（包括DNA和RNA适体和DNA酶）的滚环扩增形成的DNA和RNA超结构的性质，以扩大此类组装体作为生物分子固定化平台的用途，并将这些组装体应用于一系列生物分析装置，在这些装置中，我们将利用它们用于各种生物传感的潜力，催化和亲和分离应用。 在新的资助期内，我们将在我们正在进行的DG资助项目的基础上实现以下四个短期目标：目标1 FNA扩增子的制造：该项目的主要目标是扩大FNA扩增子NF的库，并评估NF中FNA的特性。 这项工作是该提案的基础，并将为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见解。目的2蛋白质-扩增子缀合物：该项目旨在通过杂交或适体-蛋白质相互作用将DNA寡聚体修饰的蛋白质与扩增子结合，并将这些物质用于上述应用（柱、纸传感器）。这将为我们的研究小组开辟一个全新的研究领域，并有可能在使用固定化蛋白质的生物分析中产生一些突破。目的3 DNA扩增子亲和柱：这项工作将解决一个基本问题，在使用溶胶-凝胶生物亲和柱的限制，小分子分析物。 拟议的研究将建立在我们最近的报告扩增子掺杂列，并将扩大用于形成扩增子列和这些列的整体应用程序的功能核酸的类型。目标4利用扩增子的纸质传感器：将生物分子固定到异质表面（例如纸的纤维素网络）上仍然是一个挑战。 我们将研究使用FNA扩增子作为生产多重纸传感器的平台，并将研究打印的扩增子的特性以及它们在纸表面上的行为。意义和影响：拟议的研究计划将开发一种新的方法，用于固定生物制剂（DNA适体，DNA酶和蛋白质），该方法与亲和柱，生物传感器或其他生物活性表面的制造兼容。 其结果将是一个高度通用的通用生物界面平台，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的武器库增加新工具。