CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human iPSCs
| Autor: | Amanda H. Chan, Nathaniel Huebsch, Mohammad A. Mandegar, Annie Truong, Yuichiro Miyaoka, Ekaterina B. Frolov, Po-Lin So, Tilde Eskildsen, David E. Gordon, Lei S. Qi, Luke A. Gilbert, Kristin Holmes, Søren P. Sheikh, Bruce R. Conklin, Jacqueline E. Villalta, Edward Shin, Luke M. Judge, Jonathan S. Weissman, Nevan J. Krogan, Michael P. Olvera, Max A. Horlbeck, C. Ian Spencer |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Induced Pluripotent Stem Cells Biology Regenerative Medicine Cardiovascular Medical and Health Sciences Article 03 medical and health sciences Gene expression Genetics Humans 2.1 Biological and endogenous factors Gene silencing CRISPR Clustered Regularly Interspaced Short Palindromic Repeats Gene Silencing Stem Cell Research - Embryonic - Human Aetiology Induced pluripotent stem cell Gene Psychological repression CRISPR interference Stem Cell Research - Induced Pluripotent Stem Cell Stem Cell Research - Induced Pluripotent Stem Cell - Human Cas9 Human Genome Cell Biology Biological Sciences Stem Cell Research Cell biology Heart Disease 030104 developmental biology Molecular Medicine Biotechnology Developmental Biology |
| Zdroj: | Cell stem cell, vol 18, iss 4 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, Villalta, J E, Horlbeck, M A, Gilbert, L A, Krogan, N J, Sheikh, S P, Weissman, J S, Qi, L S, So, P-L & Conklin, B R 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 |
| ISSN: | 1934-5909 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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