Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons

Aiqun Li; Samuel Cartwright; Alex Yu; Seok Man Ho; Nadine Schrode; P. J.Michael Deans; Marliette R. Matos; Meilin Fernandez Garcia; Kayla G. Townsley; Bin Zhang; Kristen J. Brennand

doi:10.1016/j.xpro.2021.100580

Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons

Aiqun Li, Samuel Cartwright, Alex Yu, Seok Man Ho, Nadine Schrode, P. J.Michael Deans, Marliette R. Matos, Meilin Fernandez Garcia, Kayla G. Townsley, Bin Zhang, Kristen J. Brennand

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

We describe a CRISPR inhibition (CRISPRi) protocol to repress endogenous gene expression (e.g., ATP6V1A) in human induced pluripotent stem cell-derived NGN2-induced glutamatergic neurons. CRISPRi enables efficient and precise gene repression of one or multiple target genes via delivering gRNA(s) to direct a dCas9-KRAB fusion protein to the gene(s) of interest. This protocol can also be adapted for gene activation and high-throughput gene manipulation, allowing assessment of the transcriptomic and phenotypic impact of candidate gene(s) associated with neurodevelopment or brain disease. For complete details on the use and execution of this protocol, please refer to Ho et al. (2017) and Wang et al. (2021).

Original language	English
Article number	100580
Journal	STAR Protocols
Volume	2
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2021.100580
State	Published - 18 Jun 2021

Keywords

CRISPR
Cell Differentiation
Molecular Biology
Neuroscience
Stem Cells

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title = "Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons",

abstract = "We describe a CRISPR inhibition (CRISPRi) protocol to repress endogenous gene expression (e.g., ATP6V1A) in human induced pluripotent stem cell-derived NGN2-induced glutamatergic neurons. CRISPRi enables efficient and precise gene repression of one or multiple target genes via delivering gRNA(s) to direct a dCas9-KRAB fusion protein to the gene(s) of interest. This protocol can also be adapted for gene activation and high-throughput gene manipulation, allowing assessment of the transcriptomic and phenotypic impact of candidate gene(s) associated with neurodevelopment or brain disease. For complete details on the use and execution of this protocol, please refer to Ho et al. (2017) and Wang et al. (2021).",

keywords = "CRISPR, Cell Differentiation, Molecular Biology, Neuroscience, Stem Cells",

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year = "2021",

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doi = "10.1016/j.xpro.2021.100580",

language = "English",

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AU - Li, Aiqun

AU - Cartwright, Samuel

AU - Yu, Alex

AU - Ho, Seok Man

AU - Schrode, Nadine

AU - Deans, P. J.Michael

AU - Matos, Marliette R.

AU - Garcia, Meilin Fernandez

AU - Townsley, Kayla G.

AU - Zhang, Bin

AU - Brennand, Kristen J.

PY - 2021/6/18

Y1 - 2021/6/18

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AB - We describe a CRISPR inhibition (CRISPRi) protocol to repress endogenous gene expression (e.g., ATP6V1A) in human induced pluripotent stem cell-derived NGN2-induced glutamatergic neurons. CRISPRi enables efficient and precise gene repression of one or multiple target genes via delivering gRNA(s) to direct a dCas9-KRAB fusion protein to the gene(s) of interest. This protocol can also be adapted for gene activation and high-throughput gene manipulation, allowing assessment of the transcriptomic and phenotypic impact of candidate gene(s) associated with neurodevelopment or brain disease. For complete details on the use and execution of this protocol, please refer to Ho et al. (2017) and Wang et al. (2021).

KW - CRISPR

KW - Cell Differentiation

KW - Molecular Biology

KW - Neuroscience

KW - Stem Cells

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ER -

Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons

Abstract

Keywords

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