CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development

Pia Annette Johansson; Anita Adami; Johan Jakobsson

doi:10.1016/j.xpro.2022.101285

CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development

Pia Annette Johansson, Anita Adami, Johan Jakobsson

Forskningsoutput: Tidskriftsbidrag › Artikel i vetenskaplig tidskrift › Peer review

Sammanfattning

This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also describe a neural progenitor differentiation protocol and discuss the challenges of target-specific lentiviral silencing, efficient silencing levels, and off-target effects. For complete details on the use and execution of this protocol, please refer to Johansson et al. (2022).

Originalspråk	engelska
Artikelnummer	101285
Tidskrift	STAR Protocols
Volym	3
Nummer	2
DOI	https://doi.org/10.1016/j.xpro.2022.101285
Status	Published - 2022 juni 17

Ämnesklassifikation (UKÄ)

Cell- och molekylärbiologi
Neurovetenskaper

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10.1016/j.xpro.2022.101285

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title = "CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development",

abstract = "This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also describe a neural progenitor differentiation protocol and discuss the challenges of target-specific lentiviral silencing, efficient silencing levels, and off-target effects. For complete details on the use and execution of this protocol, please refer to Johansson et al. (2022).",

keywords = "Cell Differentiation, CRISPR, Developmental biology, Neuroscience, Stem Cells",

author = "Johansson, {Pia Annette} and Anita Adami and Johan Jakobsson",

year = "2022",

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AB - This protocol describes the design and use of CRISPRi-mediated transcriptional silencing in human iPSCs, for loss-of-function studies in brain development research. The protocol avoids single cell selection, thereby eliminating side effects of clonal expansion and sites of viral integration. We also describe a neural progenitor differentiation protocol and discuss the challenges of target-specific lentiviral silencing, efficient silencing levels, and off-target effects. For complete details on the use and execution of this protocol, please refer to Johansson et al. (2022).

KW - Cell Differentiation

KW - CRISPR

KW - Developmental biology

KW - Neuroscience

KW - Stem Cells

U2 - 10.1016/j.xpro.2022.101285

DO - 10.1016/j.xpro.2022.101285

M3 - Article

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CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development

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Ämnesklassifikation (UKÄ)

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