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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Aug 10, 2013 |
| Title |
Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Technologies allowing for specific regulation of endogenous genes are valuable for the study of gene functions and have great potential in therapeutics. We created the CRISPR-on system, a two-component transcriptional activator consisting of a nuclease-dead Cas9 (dCas9) protein fused with a transcriptional activation domain and single guide RNAs (sgRNAs) with complementary sequence to gene promoters. We demonstrate that CRISPR-on can efficiently activate exogenous reporter genes in both human and mouse cells in a tunable manner. In addition, we show that robust reporter gene activation in vivo can be achieved by injecting the system components into mouse zygotes. Furthermore we show that CRISPR-on can activate the endogenous IL1RN, SOX2, and OCT4 genes. The most efficient gene activation was achieved by clusters of 3 to 4 sgRNAs binding to the proximal promoters suggesting their synergistic action in gene induction. Significantly, when sgRNAs targeting multiple genes were simultaneously introduced into cells, robust multiplexed endogenous gene activation was achieved. Genome-wide expression profiling demonstrated high specificity of the system.
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| Overall design |
We used microarray to assay the gene expression changes after transfection of dCas9VP160 with the different sgRNAs
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| Web link |
http://www.crispr-on.org
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| Contributor(s) |
Cheng AW, Jaenisch R |
| Citation(s) |
23979020 |
| Submission date |
Aug 09, 2013 |
| Last update date |
Dec 06, 2018 |
| Contact name |
Albert W Cheng |
| E-mail(s) |
awcheng@mit.edu
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| Organization name |
Whitehead Institute
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| Street address |
Room 453, 9 Cambridge Center
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| City |
Cambridge |
| State/province |
MA |
| ZIP/Postal code |
02139 |
| Country |
USA |
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| Platforms (1) |
| GPL571 |
[HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA214784 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE49701_RAW.tar |
5.4 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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