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

doi:10.1016/j.xpro.2021.100571

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

STAR Protoc. 2021 Jun 5;2(2):100571. doi: 10.1016/j.xpro.2021.100571. eCollection 2021 Jun 18.

Authors

Yuka A Martens^{1

2}, Siming Xu³, Richard Tait³, Gary Li³, Xinping C Zhao³, Wenyan Lu^{1

2}, Chia-Chen Liu¹, Takahisa Kanekiyo^{1

2}, Guojun Bu^{1

2}, Jing Zhao^{1

2}

Affiliations

¹ Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA.
² Neuroregeneration Lab, Center for Regenerative Medicine, Mayo Clinic, Jacksonville, FL 32224, USA.
³ ALSTEM, Richmond, CA, 94806, USA.

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).

Keywords: CRISPR; Neuroscience; Organoids; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Validation Study

MeSH terms

Brain / metabolism*
CRISPR-Cas Systems
Gene Knockdown Techniques*
Humans
Induced Pluripotent Stem Cells / metabolism*
Organoids / metabolism*

Abstract

Publication types

MeSH terms

Grants and funding