Injection and electroporation of plasmid DNA into human cortical organoids

Annina Denoth-Lippuner; Lars N Royall; Daniel Gonzalez-Bohorquez; Diana Machado; Sebastian Jessberger

doi:10.1016/j.xpro.2022.101129

Injection and electroporation of plasmid DNA into human cortical organoids

STAR Protoc. 2022 Jan 20;3(1):101129. doi: 10.1016/j.xpro.2022.101129. eCollection 2022 Mar 18.

Authors

Annina Denoth-Lippuner¹, Lars N Royall¹, Daniel Gonzalez-Bohorquez¹, Diana Machado¹, Sebastian Jessberger¹

Affiliation

¹ Laboratory of Neural Plasticity, Faculties of Medicine and Science, Brain Research Institute, University of Zurich, 8057 Zurich, Switzerland.

Abstract

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).

Keywords: Neuroscience; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cerebral Cortex / metabolism*
DNA / genetics*
Electroporation*
Humans
Organoids / metabolism*
Plasmids*
Transfection

Substances

DNA