HIGH THROUGHPUT INTRACELLULAR DELIVERY BY VISCOELASTIC MECHANOPORATION

Cell therapies such as CAR T are being explored for many disease areas but face challenges associated with safety and efficacy as well as manufacturing time and cost.Intracellular delivery of genes and gene editing complexes such as CRISPR-Cas9 are essential to manufacturing but low yield and process heterogeneity still characterize clinically available delivery methods.To address this need, we have investigated microfluidic systems that leverage the nonlinear stress behavior of viscoelastic fluids to apply submillisecond pulses of mechanical tension to the plasma membrane of cells without surface contact [1].This creates reversible pores in the membrane, allowing mRNA and CRISPR-Cas9 ribonucleoproteins to diffuse into the cytosol within seconds.This uniquely fast (>250 million cells per minute) and effective intracellular delivery platform has several promising applications in cell and gene therapy manufacturing, particularly in T cell editing for CAR T production.