DNA Nanoarrays Printed via Dip Pen Nanolithography

通过浸笔纳米光刻技术打印 DNA 纳米阵列

• 批准号: 6689822
• 负责人: Nabil Amro
• 金额: $ 10万
• 依托单位: NANOINK, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2005-03-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689822
• 关键词: fluorescent dye /probe; functional /structural genomics; glass; interdisciplinary collaboration; ionophores; microarray technology; nanotechnology; nucleic acid structure; oligonucleotides; printing; surface property; synthetic nucleotide; technology /technique development

DESCRIPTION (provided by applicant): The goal of this research is to develop novel, biologically functional DNA nanostructures that dramatically enhance the reproducibility, sensitivity, and spatial density of chip-based DNA assays. These nanostructures will improve applications ranging from point-of-care diagnosis to genomic arrays used in basic research by enabling the development of next generation screening technologies that are faster, more sensitive, more reliable, and possibly more cost effective than those presently available in the life sciences market. To accomplish the stated goals, Nanolnk will develop a DNA patterning methodology based on Dip Pen Nanolithography (DPN) to generate sub-micron sized features of DNA on solid surfaces. This multidisciplinary effort will involve life and physical scientists at Nanolnk, MEMs and instrumentation engineers at our fabrication facility, in addition to support from outside experts in the fields of DNA microarrays and microfabrication. DPN, built upon the technique of Atomic Force Microscopy (AFM), allows one to deposit materials uniformly in a direct-write fashion on surfaces with nanoscale spatial precision. This strategy offers significant advantages over current microarray printing technologies that suffer from poor spot to spot reproducibility in terms of size, shape, and oligonucleotide density, as well as reproducibility across microarray slides. Preliminary work has demonstrated that the DPN technique can be used to deposit 12mer synthetic oligonucleotides on surfaces with extremely uniform sub-100 nm to several micron scale features. The DNA nanostructures formed robust films and exhibited selectivity in binding to complementary oligonucleotides. Thus, DPN can be used to generate uniform features of synthetic DNA far smaller than can be obtained with other spotting or photolithography techniques. In Phase I, Nanolnk will demonstrate feasibility of the DPN-based approach for generating sub-micron scale DNA nanostructures on glass surfaces. The resulting nanostructures will be analyzed using existing fluorescence probe technology to provide benchmarking standards for comparison to conventional microarray assays. In addition, for applications in life sciences and biomedicine, it is desirable and advantageous in terms of speed and throughput to extend the serial patterning capability of DPN to a parallel methodology. Thus, concurrent with ink development and patterning optimization, microfabricated parallel multipen arrays will be explored as a means for faster, simultaneous writing of multiple DNA inks.

描述（由申请人提供）：本研究的目标是开发新颖的、具有生物功能的 DNA 纳米结构，显着提高基于芯片的 DNA 测定的再现性、灵敏度和空间密度。这些纳米结构将改善从现场诊断到基础研究中使用的基因组阵列的应用，通过促进下一代筛选技术的开发，这些技术比目前可用的技术更快、更灵敏、更可靠，并且可能更具成本效益。生命科学市场。为了实现既定目标，Nanolnk 将开发一种基于浸笔纳米光刻 (DPN) 的 DNA 图案化方法，以在固体表面上生成 DNA 的亚微米尺寸特征。这项多学科的努力将涉及 Nanolnk 的生命和物理科学家、我们制造工厂的 MEM 和仪器工程师，以及 DNA 微阵列和微加工领域外部专家的支持。 DPN 基于原子力显微镜 (AFM) 技术，允许人们以纳米级空间精度以直写方式在表面上均匀沉积材料。与当前的微阵列打印技术相比，该策略具有显着的优势，目前的微阵列打印技术在尺寸、形状和寡核苷酸密度以及微阵列载玻片之间的重现性方面存在较差的点与点重现性。初步工作表明，DPN 技术可用于在具有极其均匀的亚 100 nm 至几个微米尺度特征的表面上沉积 12mer 合成寡核苷酸。 DNA 纳米结构形成坚固的薄膜，并在与互补寡核苷酸结合方面表现出选择性。因此，DPN 可用于生成合成 DNA 的均匀特征，远小于使用其他点样或光刻技术获得的特征。在第一阶段，Nanolnk 将展示基于 DPN 的方法在玻璃表面生成亚微米级 DNA 纳米结构的可行性。由此产生的纳米结构将使用现有的荧光探针技术进行分析，以提供与传统微阵列分析进行比较的基准标准。此外，对于生命科学和生物医学中的应用，将DPN的串行图案化能力扩展到并行方法在速度和吞吐量方面是可取的和有利的。因此，在墨水开发和图案优化的同时，微加工并行多笔阵列将被探索作为一种更快、同时写入多种 DNA 墨水的方法。