I-Corps: Silicon Nanoneedle Chip Technology for Massively Parallel Gene Editing

I-Corps项目更广泛的影响/商业潜力将是设计一种新的基于硅的、具有成本效益的、精确的机械芯片技术，在单细胞水平上进行基因敲除和编辑。这项技术将在平行规模上实现这一目标，并最终适用于T细胞免疫疗法。利用纳米针为基础的微型机器人装置在单个细胞中定位和描绘特定基因和途径的能力将构成一项重大的技术进步，使研究人员能够监测癌症进展，研究潜在机制并开发治疗性T细胞工程。该技术主要解决与细胞和基因治疗相关的一个基本问题：在移植到人体之前用于编程细胞的病毒会引起不良的免疫反应，随着病毒DNA整合到人类基因组中，可能会造成不良反应，涉及昂贵的生产和冗长的程序，并且通常只能使用一次。这项技术将为这一基本治疗问题提供一种替代工程解决方案，可以消除这种毒性问题，包括其他领域的主要应用，包括牲畜、工业生物学、农业、药物发现和开发。I-Corps项目将进一步开发一种技术，允许编辑具有高转导效率和最小侵入性的单个T细胞。目标效率、操作和递送的敏感性/特异性、重复转导后的设备功能、不同单细胞操作的均匀性以及微机器人执行器运动的精度、再现性、滞后性和稳定性等参数将得到优化。该硅基微机器人执行器在无位置反馈传感器的开环控制下，可以在单个单元内精确跟踪和定位所需位置，从而避免了复杂的控制系统电子器件。已经开发出一个平台，其中包括一个微型机器人执行器，该执行器由一个由电容元件驱动的微平台组成，该微平台利用静电场在3D中移动，负责纳米针的独立运动。关键的创新概念是将基于并行架构的3D致动器技术与多个纳米针生物传感器集成在一起，使每个纳米针生物传感器都可以独立移动，进行有针对性的单细胞操作。

The broader impact/commercial potential of this I-Corps project will be engineering a new silicon-based cost-effective, precise, mechanical chip technology to perform gene knockdowns and editing at the single-cell level. The technology will enable this on a parallel scale as well as the eventual applicability to T cell immunotherapy. The ability to target and profile specific genes and pathways in single cells with a nanoneedle-based microrobotic device will constitute a major technological advance that will enable researchers to monitor cancer progression, study the- underlying mechanisms and develop therapeutic T cell engineering. The technology is primarily addressing a fundamental problem associated with cell and gene therapy: viruses used to program cells before transplant into the body provoke undesirable immune response, may cause adverse effects as viral DNA integrates into the human genome, involve costly production and lengthy protocols, and can typically only be used once. This technology will enable an alternative engineering solution to this fundamental therapeutic problem which can eliminate such toxicity issues encompassing major applications in other areas including livestock, industrial biology, agriculture, drug discovery and development.This I-Corps project will further develop a technology that allows editing of single T cells with high transduction efficiency and minimal invasiveness. Parameters such as target efficiency, sensitivity/specificity of manipulation and delivery, device functionality after repetitive transduction, uniformity of manipulation across different single cells, as well as precision, reproducibility, hysteresis and stability of the motion of the microrobotic actuator will be optimized. The silicon-based microrobotic actuator is designed such that it can accurately track and target desired positions within single cells under an open loop control without a position feedback sensor, thus avoiding complicated control system electronics. A platform has been developed that includes a microrobotic actuator, which consists of a microstage driven by capacitive components that moves in 3D using an electrostatic field responsible for the independent motion to the nanoneedles. The key innovative concept here is to integrate the parallel architecture based 3D actuator technology with multiple nanoneedle biosensors so that each of them can be independently moved for targeted single-cell manipulation.