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

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

Neuroscience

Research output: Contribution to journal › Article › peer-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 language	English (US)
Article number	100571
Journal	STAR Protocols
Volume	2
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2021.100571
State	Published - 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)
Medicine(all)

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10.1016/j.xpro.2021.100571

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title = "Generation and validation of APOE knockout human iPSC-derived cerebral organoids",

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",

author = "Martens, {Yuka A.} and Siming Xu and Richard Tait and Gary Li and Zhao, {Xinping C.} and Wenyan Lu and Liu, {Chia Chen} and Takahisa Kanekiyo and Guojun Bu and Jing Zhao",

note = "Funding Information: This work was supported by NIH grants R37AG027924 , RF1AG046205 , RF1AG057181 , R01AG066395 , P01NS074969 , and P30AG062677 (to G.B.), a Cure Alzheimer{\textquoteright}s Fund grant (to G.B.), and an Alzheimer{\textquoteright}s Association Research Fellowship 2018-AARF-592302 (to J.Z.). This work was also partially supported by the Mayo Clinic Center for Regenerative Medicine. Publisher Copyright: {\textcopyright} 2021 The Author(s)",

year = "2021",

month = jun,

day = "18",

doi = "10.1016/j.xpro.2021.100571",

language = "English (US)",

volume = "2",

journal = "STAR Protocols",

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TY - JOUR

T1 - Generation and validation of APOE knockout human iPSC-derived cerebral organoids

AU - Martens, Yuka A.

AU - Xu, Siming

AU - Tait, Richard

AU - Li, Gary

AU - Zhao, Xinping C.

AU - Lu, Wenyan

AU - Liu, Chia Chen

AU - Kanekiyo, Takahisa

AU - Bu, Guojun

AU - Zhao, Jing

N1 - Funding Information: This work was supported by NIH grants R37AG027924 , RF1AG046205 , RF1AG057181 , R01AG066395 , P01NS074969 , and P30AG062677 (to G.B.), a Cure Alzheimer’s Fund grant (to G.B.), and an Alzheimer’s Association Research Fellowship 2018-AARF-592302 (to J.Z.). This work was also partially supported by the Mayo Clinic Center for Regenerative Medicine. Publisher Copyright: © 2021 The Author(s)

PY - 2021/6/18

Y1 - 2021/6/18

N2 - 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).

AB - 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).

KW - CRISPR

KW - Neuroscience

KW - Organoids

KW - Stem Cells

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AN - SCOPUS:85107299124

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JO - STAR Protocols

JF - STAR Protocols

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ER -

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

Abstract

Keywords

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