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Injection and electroporation of plasmid DNA into human cortical organoids

ABSTRACT: Summary Pluripotent stem cell-derived human cortical organoids allow for the analysis of stem cell behavior and neurogenesis in neural tissues. Delivery of plasmid DNA into organoids permits visualization of individual cells, characterization of cellular components, and manipulation of gene expression. We describe a protocol to transfect cells inside organoids with plasmid DNA using micro-injection and electroporation, allowing for DNA delivery to cells within cortical units. This protocol was optimized for cortical organoids; however, it may be adapted to other organoid models. For complete details on the use and execution of this protocol, please refer to Denoth-Lippuner et al. (2021) Graphical abstract Highlights • Plasmid injection of pluripotent stem cell-derived human cortical organoids• Electroporation of human cortical organoids• Genetic manipulation of progenitors and their progeny in cortical organoids• Protocol enables robust DNA transfection of human cortical organoids Pluripotent stem cell-derived human cortical organoids allow for the analysis of stem cell behavior and neurogenesis in neural tissues. Delivery of plasmid DNA into organoids permits visualization of individual cells, characterization of cellular components, and manipulation of gene expression. We describe a protocol to transfect cells inside organoids with plasmid DNA using micro-injection and electroporation, allowing for DNA delivery to cells within cortical units. This protocol was optimized for cortical organoids; however, it may be adapted to other organoid models.

SUBMITTER: Denoth-Lippuner A

PROVIDER: S-EPMC8792447 | biostudies-literature |

REPOSITORIES: biostudies-literature

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