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

I-Corps：用于大规模并行基因编辑的硅纳米针芯片技术

• 批准号: 1740927
• 负责人: Rhonda Shrader
• 金额: $ 5万
• 依托单位: University of California-Berkeley
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1740927&HistoricalAwards=false
• 关键词: Corps; Silicon; Nanoneedle; Chip; Technology

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.

这个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成本效益，精确的机械芯片技术，以在单细胞水平上进行基因敲除和编辑。 该技术将在平行规模上实现这一点，并最终适用于T细胞免疫疗法。利用基于纳米针的微型机器人设备在单细胞中靶向和分析特定基因和途径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。该技术主要解决与细胞和基因治疗相关的一个基本问题：用于在移植到体内之前对细胞进行编程的病毒会引起不良的免疫反应，可能会导致病毒DNA整合到人类基因组中的不良反应，涉及昂贵的生产和冗长的方案，并且通常只能使用一次。该技术将为这一基本治疗问题提供替代工程解决方案，可以消除此类毒性问题，包括在其他领域的主要应用，包括畜牧业，工业生物学，农业，药物发现和开发。该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、操纵和输送的灵敏度/特异性、重复换能后的设备功能、不同单细胞间操纵的均匀性以及微型机器人致动器运动的精确度、再现性、滞后性和稳定性等参数将得到优化。硅基微机器人致动器被设计成使得其可以在开环控制下在单个单元内精确地跟踪和瞄准期望位置，而无需位置反馈传感器，从而避免复杂的控制系统电子器件。已经开发了一种平台，该平台包括微机器人致动器，该致动器由电容组件驱动的微平台组成，该电容组件使用静电场在3D中移动，该静电场负责纳米针的独立运动。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集成在一起，以便每个纳米针生物传感器都可以独立移动，以进行有针对性的单细胞操作。