Cell Stem Cell
Volume 18, Issue 4, 7 April 2016, Pages 541-553
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CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs

https://doi.org/10.1016/j.stem.2016.01.022Get rights and content
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Highlights

  • 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

Summary

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.

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Present address: Regenerative Medicine Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, 156-8506, Japan