Generation and validation of APOE knockout human iPSC-derived cerebral organoids

Yuka A. Martens, Siming Xu, Richard Tait, Gary Li, Xinping C. Zhao, Wenyan Lu, Chia-Chen Liu, Takahisa Kanekiyo, Guojun Bu, Jing Zhao

Research output: Contribution to journalArticlepeer-review

Abstract

Apolipoprotein E (apoE) is a major lipid carrier in the brain and closely associated with the pathogenesis of Alzheimer's disease (AD). Here, we describe a protocol for efficient knockout of APOE in human induced pluripotent stem cells (iPSCs) using the CRISPR-Cas9 system. We obtain homozygous APOE knockout (APOE-/-) iPSCs and further validate the deficiency of apoE in iPSC-derived cerebral organoids. APOE-/- cerebral organoids can serve as a useful tool to study apoE functions and apoE-related pathogenic mechanisms in AD. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2020).

Original languageEnglish (US)
Article number100571
JournalSTAR Protocols
Volume2
Issue number2
DOIs
StatePublished - Jun 18 2021

Keywords

  • CRISPR
  • Neuroscience
  • Organoids
  • Stem Cells

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)
  • Immunology and Microbiology(all)

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