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

University Of California-Berkeley, Berkeley CA

Investigators

Abstract

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.