CRISPRi-mediated transcriptional silencing in iPSCs for the study of human brain development
(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:
https://lup.lub.lu.se/record/4798d6fa-2cb0-4917-9935-7df7ccbed1db
- author
- Johansson, Pia Annette LU ; Adami, Anita LU and Jakobsson, Johan LU
- organization
- publishing date
- 2022-06-17
- 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
-
- scopus:85128288749
- pmid:35496780
- 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
- 2024-12-13 13:51:58
@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}}, }