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

Pia Annette Johansson; Anita Adami; Johan Jakobsson

STAR Protocols (Jun 2022)

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

Pia Annette Johansson,
Anita Adami,
Johan Jakobsson

Affiliations

Pia Annette Johansson: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden; Cell and Gene Therapy Core, Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden; Corresponding author
Anita Adami: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden
Johan Jakobsson: Laboratory of Molecular Neurogenetics, Department of Experimental Medical Science, Wallenberg Neuroscience Center and Lund Stem Cell Center, Lund University, 221 84 Lund, Sweden

Journal volume & issue: Vol. 3, no. 2
p. 101285

Abstract

Read online

Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

About the journal

Abstract

Keywords