LABEL-FREE TARGETED DELIVERY USING FEEDBACK-CONTROLLED SINGLE CELL MICRO-ELECTROPORATION IN FOCUSED ELECTRIC FIELDS

We present a device and method for continuous-flow, label-free, electronic single-cell characterization, and tandem real-time selective micro-electroporation.This enables high efficiency delivery of macromolecules to targeted cell subsets selected from heterogenous cell populations.We demonstrate simultaneous high delivery efficiency and cell viability across a range of cell types, including primary immune cell subsets from peripheral blood mononuclear cells (PBMC), and a range of delivery cargos, including CRISPR-Cas9.Scaling of cell-based therapies requires efficient, safe, and cost-effective intracellular delivery [1].Viral vectors are commonly used but significant unresolved concerns remain including high cost, payload limitations for larger cargos and safety concerns.Electroporation presents a cost-effective solution, which is payload-size agnostic [2].Conventional bulk electroporation uses high voltages (~kV) and can result in low cell viability and a trade-off with delivery efficiency for heterogenous cell populations.Electroporation is dependent on cell properties, but no singlecell level control or targeted delivery is achievable in bulk-scale methods.