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

Johansson, Pia Annette LU ; Adami, Anita LU orcid and Jakobsson, Johan LU orcid (2022) In STAR Protocols 3(2).
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).

Please use this url to cite or link to this publication:
author
; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cell Differentiation, CRISPR, Developmental biology, Neuroscience, Stem Cells
in
STAR Protocols
volume
3
issue
2
article number
101285
publisher
Cell Press
external identifiers
  • pmid:35496780
  • scopus:85128288749
ISSN
2666-1667
DOI
10.1016/j.xpro.2022.101285
language
English
LU publication?
yes
id
4798d6fa-2cb0-4917-9935-7df7ccbed1db
date added to LUP
2022-06-17 15:46:22
date last changed
2024-11-01 08:20:36
@article{4798d6fa-2cb0-4917-9935-7df7ccbed1db,
  abstract     = {{<p>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).</p>}},
  author       = {{Johansson, Pia Annette and Adami, Anita and Jakobsson, Johan}},
  issn         = {{2666-1667}},
  keywords     = {{Cell Differentiation; CRISPR; Developmental biology; Neuroscience; Stem Cells}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{2}},
  publisher    = {{Cell Press}},
  series       = {{STAR Protocols}},
  title        = {{CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development}},
  url          = {{http://dx.doi.org/10.1016/j.xpro.2022.101285}},
  doi          = {{10.1016/j.xpro.2022.101285}},
  volume       = {{3}},
  year         = {{2022}},
}