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Gene activation by dCas9-CBP and the SAM system differ in target preference
Stockholm University, Faculty of Science, Department of Molecular Biosciences, The Wenner-Gren Institute.
Stockholm University, Faculty of Science, Department of Molecular Biosciences, The Wenner-Gren Institute.
Number of Authors: 22019 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 9, article id 18104Article in journal (Refereed) Published
Abstract [en]

Gene overexpression through the targeting of transcription activation domains to regulatory DNA via catalytically defective Cas9 (dCas9) represents a powerful approach to investigate gene function as well as the mechanisms of gene control. To date, the most efficient dCas9-based activator is the Synergistic Activation Mediator (SAM) system whereby transcription activation domains are directly fused to dCas9 as well as tethered through MS2 loops engineered into the gRNA. Here, we show that dCas9 fused to the catalytic domain of the histone acetyltransferase CBP is a more potent activator than the SAM system at some loci, but less efficient at other locations in Drosophila cells. Our results suggest that different rate-limiting steps in the transcription cycle are affected by dCas9-CBP and the SAM system, and that comparing these activators may be useful for mechanistic studies of transcription as well as for increasing the number of hits in genome-wide overexpression screens.

Place, publisher, year, edition, pages
2019. Vol. 9, article id 18104
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:su:diva-177588DOI: 10.1038/s41598-019-54179-xISI: 000500551100001PubMedID: 31792240OAI: oai:DiVA.org:su-177588DiVA, id: diva2:1385729
Available from: 2020-01-15 Created: 2020-01-15 Last updated: 2020-01-15Bibliographically approved

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