High-Throughput Microfluidic Platform for Phototransfection

High throughput transient transfection of mammalian cells is crucial for intracellular gene delivery. Optoporation in a microfluidic device offers advantages over conventional physical methods for transient membrane-disruption-mediated transfection such as dosage control and scalability. Here, we describe an iterative microfluidic phototransfection platform for high throughput delivery of COVID-19 Spike Protein mRNA and CD80 mRNAs into 3T3 cells. Laminar flow focusing is used to selectively expose cells to user-defined concentrations of mRNA (dictated by scRNA seq) during which laser light is focused on individual cells to accomplish the transfection. These same cells can be repeatedly phototransfected with the same or different RNAs which is not possible with conventional approaches. This device is capable of successful phototransfection of specified numbers of RNAs into tens of thousands of cells during an experiment facilitating cell-based therapies.