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

被引：346

作者：

Mandegar, Mohammad A. ^{[1
]}

Huebsch, Nathaniel ^{[1
,2
]}

Frolov, Ekaterina B. ^{[1
]}

Shin, Edward ^{[1
]}

Truong, Annie ^{[1
]}

Olvera, Michael P. ^{[1
]}

Chan, Amanda H. ^{[1
]}

Miyaoka, Yuichiro ^{[1
,12
]}

Holmes, Kristin ^{[1
]}

Spencer, C. Ian ^{[1
]}

Judge, Luke M. ^{[1
,2
]}

Gordon, David E. ^{[3
,4
,5
]}

Eskildsen, Tilde V. ^{[6
,7
]}

Villalta, Jacqueline E. ^{[3
,4
,8
,9
]}

Horlbeck, Max A. ^{[3
,4
,8
,9
]}

Gilbert, Luke A. ^{[3
,4
,8
,9
]}

Krogan, Nevan J. ^{[3
,4
,5
]}

Sheikh, Soren P. ^{[6
,7
]}

Weissman, Jonathan S. ^{[3
,4
,8
,9
]}

Qi, Lei S. ^{[10
]}

So, Po-Lin ^{[1
]}

Conklin, Bruce R. ^{[1
,3
,4
,11
]}

机构：

[1] Gladstone Inst Cardiovasc Dis, San Francisco, CA 94158 USA

[2] Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94158 USA

[3] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94158 USA

[4] Univ Calif San Francisco, Calif Inst Quantitat Biosci, QB3, San Francisco, CA 94158 USA

[5] Gladstone Inst Virol & Immunol, San Francisco, CA 94158 USA

[6] Univ Southern Denmark, Dept Cardiovasc & Renal Res, DK-5000 Odense C, Denmark

[7] Odense Univ Hosp, Dept Clin Biochem & Pharmacol, DK-5000 Odense C, Denmark

[8] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94158 USA

[9] Univ Calif San Francisco, Ctr RNA Syst Biol, San Francisco, CA 94158 USA

[10] Stanford Univ, Dept Bioengn, Stanford, CA 94305 USA

[11] Univ Calif San Francisco, Dept Med & Cellular & Mol Pharmacol, San Francisco, CA 94158 USA

[12] Tokyo Metropolitan Inst Med Sci, Regenerat Med Project, Tokyo 1568506, Japan

来源：

CELL STEM CELL | 2016年 / 18卷 / 04期

基金：

加拿大健康研究院;

关键词：

PLURIPOTENT STEM-CELLS; TRANSCRIPTION; TALEN; DIFFERENTIATION; EXPRESSION; DISCOVERY; ACCURATE; PLATFORM; DESIGN; MOUSE;

D O I：

10.1016/j.stem.2016.01.022

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

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.

引用

页码：541 / 553

页数：13

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