The SAM complex consists of three components:
- A nucleolytically inactive Cas9-VP64 fusion
- An sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop
- The MS2-P65-HSF1 activation helper protein
The incorporation of three distinct activation domains - VP64, P65 and HSF1 – into the complex aids robust transcriptional activation through synergy.
SAM is capable of robust activation of coding as well as non-coding elements (lincRNA). For targeted activation of coding genes, please look at our activator sgRNA design tool.
SAM can further be combined with a human genome-wide library to activate all known coding isoforms form the RefSeq database (23,430 isoforms) for gain-of-function screening.See: Konermann S*, Brigham MD*, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O & Zhang F. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature, doi:10.1038/nature14136.
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