Raab Matthew, Hancock William O
Department of Bioengineering, 229 Hallowell Building, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
Biotechnol Bioeng. 2008 Mar 1;99(4):764-73. doi: 10.1002/bit.21645.
Shrinking biosensors down to microscale dimensions enables increases in sensitivity and the ability to analyze minute samples such as the contents of individual cells. The goal of the present study is to create mobile microscale biosensors by attaching molecular beacons to microtubules and using kinesin molecular motors to transport these functionalized microtubules across two-dimensional surfaces. Previous work has shown that microfluidic channels can be functionalized with kinesin motors such that microtubules can be transported and directed through these channels without the need for external power or pressure-driven pumping. In this work, we show that molecular beacons can be attached to microtubules such that both the fluorescence reporting capability of the beacon and the motility of the microtubules are retained. These molecular beacon-functionalized microtubules were able to bind ssDNA target sequences, transport them across surfaces, and report their presence by an increase in fluorescence that was detected by fluorescence microscopy. This work is an important step toward creating hybrid microdevices for sensitive virus detection or analyzing mRNA profiles of individual cells.
将生物传感器缩小到微尺度尺寸能够提高灵敏度，并增强分析微量样品（如单个细胞内容物）的能力。本研究的目标是通过将分子信标附着到微管上，并利用驱动蛋白分子马达在二维表面上运输这些功能化微管，来制造可移动的微尺度生物传感器。先前的工作表明，微流体通道可以用驱动蛋白马达进行功能化，从而使微管能够在无需外部电源或压力驱动泵的情况下在这些通道中运输和定向。在这项工作中，我们表明分子信标可以附着到微管上，从而使信标的荧光报告能力和微管的运动性都得以保留。这些分子信标功能化的微管能够结合单链DNA靶序列，在表面上运输它们，并通过荧光显微镜检测到的荧光增强来报告它们的存在。这项工作是朝着制造用于灵敏病毒检测或分析单个细胞mRNA谱的混合微器件迈出的重要一步。
