DNA Nanoarrays Printed via Dip Pen Nanolithography

描述(申请人提供)：这项研究的目标是开发新型的、具有生物功能的DNA纳米结构，显著提高基于芯片的DNA分析的重复性、灵敏度和空间密度。这些纳米结构将通过开发比目前生命科学市场上可用的技术更快、更灵敏、更可靠、可能更具成本效益的下一代筛查技术，来改进从护理点诊断到基础研究中使用的基因组阵列的各种应用。为了实现上述目标，Nanolnk将开发一种基于浸笔纳米光刻(DPN)的DNA图案化方法，以在固体表面生成亚微米尺寸的DNA特征。这一多学科的努力将涉及Nanolnk的生命和物理学家、MEMS和我们制造设施的仪器工程师，以及DNA微阵列和微制造领域的外部专家的支持。DPN建立在原子力显微镜(AFM)技术的基础上，可以在纳米级的空间精度的表面上以直接写入的方式均匀地沉积材料。与当前的微阵列打印技术相比，该策略提供了显著的优势，目前的微阵列打印技术在大小、形状和寡核苷酸密度以及跨微阵列载玻片的重复性方面存在点对点重现性差的问题。初步工作表明，DPN技术可以用来在具有极均匀的亚100 nm到几个微米级特征的表面沉积12聚体合成寡核苷酸。DNA纳米结构形成了坚固的薄膜，并显示出与互补寡核苷酸结合的选择性。因此，DPN可以用来产生合成DNA的均匀特征，远远小于其他斑点或光刻技术所能获得的特征。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，以提供基准标准，以与传统的微阵列分析进行比较。此外，对于生命科学和生物医学中的应用，将DPN的串联构图能力扩展到并行方法在速度和吞吐量方面是理想的和有利的。因此，在墨水开发和图案化优化的同时，微细制造的并行多笔阵列将被探索为一种更快地同时写入多个DNA墨水的手段。

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.