Nanomechanical biosensing with aptazymes selected for surface function

本提案的目的是为基于纳米机械悬臂梁的生物传感器开发新的生物分子信号放大方案。该方法是选择适体酶（基于RNA的酶分子），其基于其在传感器表面上起作用的能力执行靶激活连接，并通过采用滚环扩增（RCA）方案显著增强悬臂弯曲信号。该项目的具体目标是：1）基于溶液的适体酶和RCA适应悬臂梁，2）开发新的表面功能适体酶和3）基于表面的适体酶适应纳米机械悬臂梁检测蛋白质。在本研究中，Savran博士将担任PI，负责所有实验。Ellington博士将担任共同主要研究员，负责分子选择和合成。智力优势：在这项研究中，将首次展示用于传感器表面最佳性能的适体酶的合成。所选择的分子将适应纳米机械悬臂梁传感器沿着滚环放大，以实现仅通过基于荧光的检测系统观察到的增强的信号和检测限。广泛影响：该项目将通过微加工技术大规模生产超灵敏探测器，降低单位设备的成本。预计这将产生重大的经济和社会影响，使负担得起的系统，以各种各样的用户，从生物学家到临床医生。该项目还旨在i）促进高中学生的纳米技术意识，ii）促进少数民族背景的本科生参与纳米技术研究。

The objective of this proposal is to develop novel biomolecular signal amplification protocols for nanomechanical cantilever based biosensors. The approach is to select aptazymes (RNA-based enzymatic molecules) that perform target-activated ligation based their ability to function on sensor surfaces and to enhance cantilever bending signals significantly by employing rolling circle amplification (RCA) protocols. The specific aims of the project are: 1) Adaptation of solution-based aptazymes and RCA to the cantilever, 2) Development of new aptazymes for surface function and 3) Adaptation of surface-based aptazymes to the nanomechanical cantilever for detection of proteins. In this study, Dr. Savran will be the PI and will be responsible for all experiments. Dr. Ellington will be co-PI and will undertake molecular selection and synthesis.Intellectual Merit: In this study, the synthesis of aptazymes for optimal performance on sensor surfaces will be demonstrated for the first time. The selected molecules will be adapted to the nanomechanical cantilever sensor along with rolling circle amplification to achieve enhanced signals and detection limits that are only observed by fluorescence-based detection systems. Broad Impact: This project will result in ultrasensitive detectors that are mass produced via microfabrication techniques with reduced costs per unit device. This is expected to have significant economical and social impacts by enabling affordable systems to a wide variety of users ranging from biologists to clinicians. This project will also aim to i) contribute to the nanotechnology awareness of high-school students and ii) foster involvement of undergraduate students of minority backgrounds in nanotechnology research.