Abstract：
: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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Sajwan S,Mannervik Mdoi
10.1038/s41598-019-54179-xsubject
Has Abstractpub_date
2019-12-02 00:00:00pages
18104issue
1issn
2045-2322pii
10.1038/s41598-019-54179-xjournal_volume
9pub_type
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