DNA Nanoarrays Printed via Dip Pen Nanolithography

描述（由申请人提供）：本研究的目标是开发新颖的，具有生物学功能的DNA纳米结构，以显着提高基于芯片的DNA分析的可重复性，灵敏度和空间密度。这些纳米结构将改善从即时诊断到用于基础研究的基因组阵列的应用，使下一代筛查技术的发展更快、更敏感、更可靠，并且可能比目前生命科学市场上现有的筛查技术更具成本效益。为了实现既定的目标，nanolink将开发一种基于蘸笔纳米光刻（DPN）的DNA图谱方法，以在固体表面上生成亚微米尺寸的DNA特征。除了DNA微阵列和微制造领域的外部专家的支持外，这项多学科的努力将涉及nanolink的生命和物理科学家，我们制造工厂的MEMs和仪器工程师。DPN建立在原子力显微镜（AFM）技术的基础上，允许人们以纳米级空间精度的方式均匀地将材料沉积在表面上。与当前的微阵列打印技术相比，这种策略具有显著的优势。当前的微阵列打印技术在尺寸、形状、寡核苷酸密度以及微阵列载玻片的可重复性方面存在点到点的可重复性差。初步研究表明，DPN技术可用于在亚100纳米到几微米尺度的表面上沉积12聚合成寡核苷酸。DNA纳米结构形成了坚固的薄膜，并表现出与互补寡核苷酸结合的选择性。因此，DPN可用于生成合成DNA的均匀特征，远小于其他斑点或光刻技术所能获得的特征。在第一阶段，nanolink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。所得到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比较的基准标准。此外，对于生命科学和生物医学的应用，将DPN的串行模式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，随着墨水的发展和图案的优化，微制造并行多笔阵列将被探索作为一种更快，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.