Human CRISPR Activation Library (SAM - 3 plasmid system)
(Pooled Library #1000000057, #1000000074)
The human CRISPR/Cas9 Synergistic Activation Mediator (SAM) pooled libraries use an engineered protein complex for the transcriptional activation of endogenous genes.
This library consists of three components which are all provided:
- A nucleolytically inactive Cas9-VP64 fusion (Addgene plasmid # 61425)
- A gRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2 (present in the libraries)
- The MS2-P65-HSF1 plasmid which expresses the activation helper protein (Addgene plasmid #89308)
Note: Libraries ordered prior to 4/3/2017 were shipped with Addgene plasmid #61426 rather than #89308
- Depositing Labs
Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.
You will also receive bacterial stabs of the MS2-P65-HSF1 plasmid (Addgene #89308) and dCas9-VP64 (Addgene #61425). Please do NOT order these plasmids in addition to this library. These plasmids are SHIPPED SEPARATELY at ambient temperature.
Terms and Licenses
The SAM library consists of 3 unique sgRNAs targeting the RefSeq coding isoforms in the proximal promoter (>90% of sgRNAs are targeted to the first 200bp upstream of the transcription start site of their target).
The Zeocin resistant gRNA pooled library is referred to as #61597 in Konermann et al, 2014.
For additional information, protocols and an activator sgRNA design tool, visit the Zhang Lab SAM website: http://sam.genome-engineering.org/
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the pooled libraries were described, and include Addgene in the Materials and Methods of your future publications.
Example for your Materials & Methods section:Human CRISPR 3-plasmid activation pooled library (SAM) was a gift from Feng Zhang (Addgene # )
For your References section:Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S*, Brigham MD*, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O & Zhang F. Nature 2015 Jan 29;517(7536):583-8. doi: 10.1038/nbt.3437. 10.1038/nature14136. Epub 2014 Dec 10PubMed 25494202