(1179-D) Comparison of Transient Gene Delivery Methods in Human iPSC-derived Neurons and Glia
Tuesday, February 6, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: Human induced pluripotent stem cell (hiPSC)-derived brain cells provide valuable platforms to study underlying molecular mechanisms of human neurological disorders. These studies often require genetic manipulation of molecular pathways, through protein overexpression and gene knockdown, to identify gene targets for therapeutic development. It is well-known that delivery of synthetic nucleic acids into brain cells is challenging and often results in low transfection efficiencies and low cell survival rates after transfection. While numerous research studies have focused on transient gene delivery to primary neurons isolated from rodent models, similar studies for human iPSC-derived brain cells have been limited. Therefore, we aimed to determine optimal transfection methods for hiPSC-derived brain cells that would yield high transfection efficiency and low cytotoxicity. We performed chemical (cationic lipid-based), physical (electroporation), and biological (viral delivery) transfection methods to deliver a fluorescent reporter gene that is expressed under a ubiquitous promoter. Our results showed that biological transfection methods provide the highest transfection efficiency with at least a week of cell survival for both hiPSC-derived neurons and glia, while chemical transfection methods result in the lowest transfection efficiency with the highest toxicity. This study provides guidance for researchers who aim to overexpress or knockdown genes of interest transiently in relevant human brain cells to understand their functions and identify therapeutic targets in neurological disorders.