Discovery of an autoimmunity-associated IL2RA enhancer by unbiased targeting of transcriptional activation
| Autor: | Eric Boyer, Mandy Boontanrart, Mark J. Daly, Alice Y. Chan, Nicolas Bray, Theodore L. Roth, Frédéric Van Gool, Jonathan M. Woo, Dimitre R. Simeonov, Mark S. Anderson, Kyle K. Fahr, Gemma L. Curie, Therese Mitros, Jacob E. Corn, Alexander Marson, Meena Subramaniam, Nicki Naddaf, Kathrin Schumann, Chun J Ye, Jeffrey A. Bluestone, Youjin Lee, Graham J. Ray, Michelle L.T. Nguyen, Rachel E. Gate, Benjamin G. Gowen |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Genetics
Regulation of gene expression 0303 health sciences Enhancer RNAs Biology medicine.disease_cause Autoimmunity 03 medical and health sciences 0302 clinical medicine Genome editing Genetic variation medicine CRISPR Enhancer Transcription factor 030217 neurology & neurosurgery 030304 developmental biology |
| DOI: | 10.1101/091843 |
| Popis: | The majority of genetic variants associated with common human diseases map to enhancers, non-coding elements that shape cell type-specific transcriptional programs and responses to specific extracellular cues 1-3. In order to understand the mechanisms by which non-coding genetic variation contributes to disease, systematic mapping of functional enhancers and their biological contexts is required. Here, we develop an unbiased discovery platform that can identify enhancers for a target gene without prior knowledge of their native functional context. We used tiled CRISPR activation (CRISPRa) to synthetically recruit transcription factors to sites across large genomic regions (>100 kilobases) surrounding two key autoimmunity risk loci, CD69 and IL2RA (interleukin-2 receptor alpha; CD25). We identified several CRISPRa responsive elements (CaREs) with stimulation-dependent enhancer activity, including an IL2RA enhancer that harbors an autoimmunity risk variant. Using engineered mouse models and genome editing of human primary T cells, we found that sequence perturbation of the disease-associated IL2RA enhancer does not block IL2RA expression, but rather delays the timing of gene activation in response to specific extracellular signals. This work develops an approach to rapidly identify functional enhancers within non-coding regions, decodes a key human autoimmunity association, and suggests a general mechanism by which genetic variation can cause immune dysfunction. |
| Databáze: | OpenAIRE |
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