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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.
Affiliation
  • Li A; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Cartwright S; Mount Sinai Center for Transformative Disease Modeling, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Yu A; Icahn Institute for Data Science and Genomic Technology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Ho SM; Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, New York, NY 10029, USA.
  • Schrode N; Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, New York, NY 10029, USA.
  • Deans PJM; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Matos MR; Mount Sinai Center for Transformative Disease Modeling, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Garcia MF; Icahn Institute for Data Science and Genomic Technology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Townsley KG; Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Zhang B; Mount Sinai Center for Transformative Disease Modeling, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
  • Brennand KJ; Icahn Institute for Data Science and Genomic Technology, Icahn School of Medicine at Mount Sinai, One Gustave L. Levy Place, New York, NY 10029, USA.
STAR Protoc ; 2(2): 100580, 2021 06 18.
Article in En | MEDLINE | ID: mdl-34151300
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).
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression Regulation / Induced Pluripotent Stem Cells / CRISPR-Cas Systems / Nerve Tissue Proteins / Neurons Limits: Humans Language: En Journal: STAR Protoc Year: 2021 Type: Article Affiliation country: United States
Fulltext
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression Regulation / Induced Pluripotent Stem Cells / CRISPR-Cas Systems / Nerve Tissue Proteins / Neurons Limits: Humans Language: En Journal: STAR Protoc Year: 2021 Type: Article Affiliation country: United States