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).
| Original language | English |
|---|---|
| Article number | 101285 |
| Journal | STAR Protocols |
| Volume | 3 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2022 Jun 17 |
Subject classification (UKÄ)
- Cell and Molecular Biology
- Neurosciences
Free keywords
- Cell Differentiation
- CRISPR
- Developmental biology
- Neuroscience
- Stem Cells