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

这个i-Corps项目的更广泛的影响/商业潜力将是设计一种新的基于硅的成本效益高、精确的机械芯片技术，以在单细胞水平上执行基因敲除和编辑。这项技术将在平行规模上实现这一点，并最终应用于T细胞免疫治疗。利用基于纳米针的微型机器人设备在单细胞中靶向和描绘特定基因和通路的能力将构成一项重大技术进步，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。这项技术主要是解决与细胞和基因治疗相关的一个根本问题：在移植到体内之前对细胞进行编程的病毒会引发不良的免疫反应，随着病毒DNA整合到人类基因组中，可能会产生不良影响，涉及昂贵的生产和冗长的程序，而且通常只能使用一次。这项技术将为这一根本的治疗问题提供另一种工程解决方案，可以消除这种毒性问题，包括在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，允许以高转导效率和最小的侵入性编辑单个T细胞。目标效率、操作和递送的灵敏度/特异性、重复转导后的设备功能、跨不同单细胞的操作的一致性以及微机器人执行器运动的精确度、重复性、滞后性和稳定性等参数将被优化。硅基微机器人执行器的设计使得它可以在开环控制下精确跟踪和定位单个电池内的所需位置，而不需要位置反馈传感器，从而避免了复杂的控制系统电子设备。已经开发出一种包括微机器人致动器的平台，该微机器人致动器由由电容组件驱动的微工作台组成，该微工作台利用负责向纳米针独立运动的静电场在3D中移动。这里的关键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相结合，以便每个传感器都可以独立移动，以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.