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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).

SUBMITTER: Johansson PA

PROVIDER: S-EPMC9038555 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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

Johansson Pia Annette PA Adami Anita A Jakobsson Johan J

STAR protocols 20220413 2

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 t ...[more]

PMID: 35496780

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