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

Mohammad A. Mandegar; Nathaniel Huebsch; Ekaterina B. Frolov; Edward Shin; Annie Truong; Michael P. Olvera; Amanda H. Chan; Yuichiro Miyaoka; Kristin Holmes; C. Ian Spencer; Luke M. Judge; David E. Gordon; Tilde V. Eskildsen; Jacqueline E. Villalta; Max A. Horlbeck; Luke A. Gilbert; Nevan J. Krogan; Søren P. Sheikh; Jonathan S. Weissman; Lei S. Qi; Po Lin So; Bruce R. Conklin

doi:10.1016/j.stem.2016.01.022

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

Mohammad A. Mandegar, Nathaniel Huebsch, Ekaterina B. Frolov, Edward Shin, Annie Truong, Michael P. Olvera, Amanda H. Chan, Yuichiro Miyaoka, Kristin Holmes, C. Ian Spencer, Luke M. Judge, David E. Gordon, Tilde V. Eskildsen, Jacqueline E. Villalta, Max A. Horlbeck, Luke A. Gilbert, Nevan J. Krogan, Søren P. Sheikh, Jonathan S. Weissman, Lei S. QiPo Lin So, Bruce R. Conklin

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Abstract

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.

Original language	English
Pages (from-to)	541-553
Number of pages	13
Journal	Cell Stem Cell
Volume	18
Issue number	4
DOIs	https://doi.org/10.1016/j.stem.2016.01.022
State	Published - 7 Apr 2016
Externally published	Yes

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Mandegar, M. A., Huebsch, N., Frolov, E. B., Shin, E., Truong, A., Olvera, M. P., Chan, A. H., Miyaoka, Y., Holmes, K., Spencer, C. I., Judge, L. M., Gordon, D. E., Eskildsen, T. V., Villalta, J. E., Horlbeck, M. A., Gilbert, L. A., Krogan, N. J., Sheikh, S. P., Weissman, J. S., ... Conklin, B. R. (2016). CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs. Cell Stem Cell, 18(4), 541-553. https://doi.org/10.1016/j.stem.2016.01.022

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title = "CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs",

abstract = "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.",

author = "Mandegar, {Mohammad A.} and Nathaniel Huebsch and Frolov, {Ekaterina B.} and Edward Shin and Annie Truong and Olvera, {Michael P.} and Chan, {Amanda H.} and Yuichiro Miyaoka and Kristin Holmes and Spencer, {C. Ian} and Judge, {Luke M.} and Gordon, {David E.} and Eskildsen, {Tilde V.} and Villalta, {Jacqueline E.} and Horlbeck, {Max A.} and Gilbert, {Luke A.} and Krogan, {Nevan J.} and Sheikh, {S{\o}ren P.} and Weissman, {Jonathan S.} and Qi, {Lei S.} and So, {Po Lin} and Conklin, {Bruce R.}",

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year = "2016",

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Mandegar, MA, Huebsch, N, Frolov, EB, Shin, E, Truong, A, Olvera, MP, Chan, AH, Miyaoka, Y, Holmes, K, Spencer, CI, Judge, LM, Gordon, DE, Eskildsen, TV, Villalta, JE, Horlbeck, MA, Gilbert, LA, Krogan, NJ, Sheikh, SP, Weissman, JS, Qi, LS, So, PL & Conklin, BR 2016, 'CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs', Cell Stem Cell, vol. 18, no. 4, pp. 541-553. https://doi.org/10.1016/j.stem.2016.01.022

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AU - Mandegar, Mohammad A.

AU - Huebsch, Nathaniel

AU - Frolov, Ekaterina B.

AU - Shin, Edward

AU - Truong, Annie

AU - Olvera, Michael P.

AU - Chan, Amanda H.

AU - Miyaoka, Yuichiro

AU - Holmes, Kristin

AU - Spencer, C. Ian

AU - Judge, Luke M.

AU - Gordon, David E.

AU - Eskildsen, Tilde V.

AU - Villalta, Jacqueline E.

AU - Horlbeck, Max A.

AU - Gilbert, Luke A.

AU - Krogan, Nevan J.

AU - Sheikh, Søren P.

AU - Weissman, Jonathan S.

AU - Qi, Lei S.

AU - So, Po Lin

AU - Conklin, Bruce R.

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AB - 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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JO - Cell Stem Cell

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

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