URoL:ASC:Synthetic Protocell Communities to Address Critical Sensing Challenges

该项目将创建一种可供未经培训的用户使用的工具，无需复杂的设备，即可诊断以多个指示剂分子组合为特征的医疗状况。虽然一些疾病可以通过单个分子的水平来诊断，但大多数需要测量多个分子的水平才能进行准确的诊断。为了以用户友好的方式应对这一挑战，该项目需要创建一种工具，利用生物学来感知目标分子，结合从这些测量中获得的信息，并向用户报告患者状态。该项目使用了一种技术，可以在模仿简化版细胞的小液滴中开启和关闭基因。单个液滴感知单个分子，然后与另一个液滴通信，产生联合读数。通过让每个液滴检测单个分子并利用与其他液滴的可重复使用的通信形式，它使该工具成为模块化的，基本上可以为任何一组目标分子创建诊断。液滴技术以及所提出的信号通信和组合方法的组合是独特和新颖的。如果成功完成，该项目将对社会产生广泛影响，并通过能够评估和监测发展中国家和世界各地的营养状况、帮助个人和改善公共卫生政策，帮助解决健康和其他不平等问题。作为原则试验的证明，该项目侧重于开发锌缺乏症的综合诊断方法，锌缺乏症每年导致数万名5岁以下儿童死亡。液滴技术使用双水相系统和无细胞转录和翻译相结合的方式来创建“原细胞”，从而实现用于传感和信号集成的生物电路，从而创建复杂的生物传感器。多个原细胞各自产生一种不同的信使分子，从自然界中提取，以实现与单独的“接收器”原细胞的通信，该原细胞通过一个三杂交系统整合信号。通过使用通用的通信和集成框架，单个传感器原细胞可以改变为感知不同的目标分子，而不会影响下游的生物计算，这大大简化了针对不同条件创建新的诊断方法。该项目的一个重要部分还将包括努力使Protocell平台更有效率和更强大，以供现场使用。这些努力将构建合成细胞和微生物组相互作用和机制的生命规则作为开发平台的基础，该平台可以更容易地获得医疗诊断，从而对营养流行病学和患者健康产生重大影响。该项目还设想并将涉及对拟议平台的环境应用。这些努力，加上外展努力和对工程学中代表性不足群体的学生的支持，预计将产生真正广泛的影响。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project will create a tool that can be used by untrained users and without sophisticated equipment to allow diagnosis of medical conditions characterized by the combination of multiple indicator molecules. While some medical conditions can be diagnosed by the level of just a single molecule, most require the measurement of multiple molecules’ levels to enable accurate diagnosis. To address this challenge in a user-friendly manner, this project entails creating a tool that harnesses biology to sense target molecules, combine the information gained from those measurements, and report out patient status to the user. The project uses technology that allows genes to be turned on and off in small droplets that mimic simplified versions of cells. Individual droplets sense an individual molecule and then communicate with another droplet that produces a combined readout. By having each droplet detect a single molecule and exploit a reusable form of communication with other droplets, it makes the tool modular for creation of diagnostics for essentially any set of target molecules. The droplet technology and the combination of proposed approaches for communication and combination of signals are unique and novel. If successfully completed, this project would impact society broadly and help address health and other inequities by enabling assessment and monitoring of nutrition status in developing countries and around the world, helping individuals and improving public health policy.As a proof of principle testbed, the project focuses on the development of a comprehensive diagnostic for zinc deficiency, a condition that is responsible for the deaths of tens of thousands of children under the age of 5 every year. The droplet technology uses aqueous two-phase systems combined with cell-free transcription and translation to create “protocells” that enable the implementation of biological circuits for sensing and signal integration and thus the creation of complex biosensors. Multiple protocells each produce a distinct messenger molecule drawn from nature to enable communication with a separate “receiver” protocell that integrates the signal via a three-hybrid system. By using a common communication and integration framework, individual sensor protocells can be changed to sense different target molecules without impacting the downstream biological computation, which greatly simplifies the creation of new diagnostics for different conditions. An important part of the project will also include efforts to make the protocell platform more efficient and robust for field use. These efforts use the Rules of Life for Building a Synthetic Cell and for Microbiome Interactions and Mechanisms as the basis for the development of a platform that could have a substantial impact on nutritional epidemiology and patient health by virtue of easier access to medical diagnostics. The project also envisions and will entail pursuit of environmental applications of the proposed platform. These efforts, combined with outreach efforts and support for students from groups that are underrepresented in engineering, are expected to have a truly broad impact.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.