CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs

Developing technologies for efficient and scalable disruption of gene expression will provide powerful tools for studying gene function, developmental pathways, and disease mechanisms. Here, we develop clustered regularly interspaced short palindromic repeat interference (CRISPRi) to repress gene expression in human induced pluripotent stem cells (iPSCs). CRISPRi, in which a doxycycline-inducible deactivated Cas9 is fused to a KRAB repression domain, can specifically and reversibly inhibit gene expression in iPSCs and iPSC-derived cardiac progenitors, cardiomyocytes, and T lymphocytes. This gene repression system is tunable and has the potential to silence single alleles. Compared with CRISPR nuclease (CRISPRn), CRISPRi gene repression is more efficient and homogenous across cell populations. The CRISPRi system in iPSCs provides a powerful platform to perform genome-scale screens in a wide range of iPSC-derived cell types, dissect developmental pathways, and model disease. [Display omitted] •Inducible CRISPRi iPSCs provide a valuable resource for rapid gene knockdown•CRISPRi knockdown is efficient, tunable, and reversible in iPSCs•CRISPRi knockdown is highly specific•CRISPRi enables disease modeling in iPSC-derived cardiomyocytes In this article, Mandegar and colleagues utilize CRISPR interference for efficient gene knockdown in iPSCs and their differentiated cell derivatives. The CRISPRi tools and cell lines presented in this study are highly versatile and serve as a useful resource for the cell and stem cell biology communities.