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

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...

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Autores principales: Li, Aiqun, Cartwright, Samuel, Yu, Alex, Ho, Seok-Man, Schrode, Nadine, Deans, P.J. Michael, Matos, Marliette R., Garcia, Meilin Fernandez, Townsley, Kayla G., Zhang, Bin, Brennand, Kristen J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8188621/
https://www.ncbi.nlm.nih.gov/pubmed/34151300
http://dx.doi.org/10.1016/j.xpro.2021.100580
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author Li, Aiqun
Cartwright, Samuel
Yu, Alex
Ho, Seok-Man
Schrode, Nadine
Deans, P.J. Michael
Matos, Marliette R.
Garcia, Meilin Fernandez
Townsley, Kayla G.
Zhang, Bin
Brennand, Kristen J.
author_facet Li, Aiqun
Cartwright, Samuel
Yu, Alex
Ho, Seok-Man
Schrode, Nadine
Deans, P.J. Michael
Matos, Marliette R.
Garcia, Meilin Fernandez
Townsley, Kayla G.
Zhang, Bin
Brennand, Kristen J.
author_sort Li, Aiqun
collection PubMed
description 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).
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spelling pubmed-81886212021-06-17 Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons Li, Aiqun Cartwright, Samuel Yu, Alex Ho, Seok-Man Schrode, Nadine Deans, P.J. Michael Matos, Marliette R. Garcia, Meilin Fernandez Townsley, Kayla G. Zhang, Bin Brennand, Kristen J. STAR Protoc Protocol 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). Elsevier 2021-06-03 /pmc/articles/PMC8188621/ /pubmed/34151300 http://dx.doi.org/10.1016/j.xpro.2021.100580 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Li, Aiqun
Cartwright, Samuel
Yu, Alex
Ho, Seok-Man
Schrode, Nadine
Deans, P.J. Michael
Matos, Marliette R.
Garcia, Meilin Fernandez
Townsley, Kayla G.
Zhang, Bin
Brennand, Kristen J.
Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title_full Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title_fullStr Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title_full_unstemmed Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title_short Using the dCas9-KRAB system to repress gene expression in hiPSC-derived NGN2 neurons
title_sort using the dcas9-krab system to repress gene expression in hipsc-derived ngn2 neurons
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8188621/
https://www.ncbi.nlm.nih.gov/pubmed/34151300
http://dx.doi.org/10.1016/j.xpro.2021.100580
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