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

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...

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Bibliographic Details
Published in:STAR protocols 2021-06, Vol.2 (2), p.100571-100571, Article 100571
Main Authors: Martens, Yuka A., Xu, Siming, Tait, Richard, Li, Gary, Zhao, Xinping C., Lu, Wenyan, Liu, Chia-Chen, Kanekiyo, Takahisa, Bu, Guojun, Zhao, Jing
Format: Article
Language:English
Subjects:
Brain - metabolism
CRISPR
CRISPR-Cas Systems
Gene Knockdown Techniques
Humans
Induced Pluripotent Stem Cells - metabolism
Neuroscience
Organoids
Organoids - metabolism
Protocol
Stem Cells
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Summary: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). [Display omitted] •Detailed protocol to generate APOE knockout human iPSC lines via CRISPR-Cas9 technology•Detailed protocol to generate cerebral organoids from parental and isogenic iPSC lines•Steps for quality control and assessment of apoE deletion in cerebral organoids•Provides a valuable tool for apoE function study using iPSC-derived cerebral organoids 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.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2021.100571