Rolosense: An innovative platform for automatic mobile phone readout of active SARS-CoV-2 particles

项目摘要 本方案的最终目标是开发一种新型的自动监控平台技术 以及真实的追踪空气中的SARS-CoV-2病毒颗粒。该提案的核心是 “Rolosense”技术利用DNA微马达作为病毒传感和转导 材料（VSTM），其可由常规智能手机相机检测。这既提供 地理追踪和监视。Rolosense马达由DNA包覆的球形颗粒组成 （5微米直径），其与用互补RNA修饰的表面杂交。颗粒 在加入RNase H后以超过1微米/分钟的速度移动，RNase H选择性地水解 杂交RNA，而不是单链RNA。病毒结合配体（VBL）包被的DNA马达失速 在存在SARS-CoV-2病毒颗粒的情况下。因为马达可以自动移动， 在没有干预的情况下，该测定是完全自动化的，并且常规步骤，如病毒检测， 不需要灭活、RNA分离和扩增。读出是使用 自动化智能手机应用程序，无需分光光度计或 荧光计。初步数据显示了实时SARS-CoV-2假病毒颗粒传感。里程碑 包括高亲和力和高特异性VBL的筛选和鉴定。适体和 将筛选和验证抗体VBL。模拟和实验将被用来理解 温度和环境条件在调节Rolosense性能中的作用。 VBL与DNA折纸的多价展示将增强亲合力。最后，微流控芯片， 将实施和测试空中飞沫捕捉。这项工作将由一个高度 这是一个跨学科团队，具有互补的专门知识和共同出版物的记录。PI Salaita 发明了Rolosense技术，并在开发手机诊断方面拥有丰富的经验， 合成马达Co-I Melikian是病毒学家，Co-I Heemstra是开发 作为新型靶点的适体，Co-I Ke在DNA折纸结构方面具有丰富的经验， 绑定，并共同撰写了工作的Rolosense，Co-I Rajaraman和Primordia是专家， 微流体装置的开发和商业化。我们的解决方案提供了一种潜力， 立即解决当今迫切的病毒检测和跟踪需求。

Project Abstract The ultimate goal of this proposal is to develop a novel platform technology for automatic surveillance and tracing of airborne SARS-CoV-2 virus particles in real time. The centerpiece of this proposal is the “Rolosense” technology which leverages a DNA micromotor as the virus sensing and transduction material (VSTM) that can be detected by a conventional smart phone camera. This provides both geographical tracing and surveillance. Rolosense motor are comprised a DNA-coated spherical particle (5 micrometer diameter) that hybridizes to a surface modified with complementary RNA. The particle moves at speeds of over 1 micron/minute upon the addition of RNase H, which selectively hydrolyses hybridized RNA but not single-stranded RNA. DNA motors coated with virus binding ligand (VBL) stall in presence of SARS-CoV-2 virus particles. Because motors move autonomously for distances up to millimeters without intervention, the assay is fully automated, and conventional steps such as viral inactivation, RNA isolation and amplification are not required. The readout is performed using an automated smart phone app for particle tracking without the need for a spectrophotometer or fluorometer. Preliminary data shows realtime SARS-CoV-2 pseudovirus particle sensing. Milestones include the screening and identification of high affinity and high specificity VBLs. Both aptamers and antibody VBLs will be screening and validated. Simulations and experiments will be used to understand the role of temperature and environmental conditions in modulating Rolosense performance. Multivalent display of VBLs with DNA origami will enhance avidity. Finally, microfluidic chips with airborne droplet capture will be implemented and tested. The work will be performed by a highly interdisciplinary team with complementary expertise and a track-record of co-publications. PI Salaita invented the Rolosense technology and has past experience in developing cell phone diagnostics and synthetic motors. Co-I Melikian is an expert virologist, Co-I Heemstra is an expert at developing aptamers for novel targets, Co-I Ke has extensive experience in DNA origami structures for avid target binding and has co-authored work on Rolosense, Co-I Rajaraman and Primordia are experts at microfluidic device development and commercialization. Our solution offers the potential to provide an immediate solution to today’s urgent virus sensing and tracing needs.