SAM Pooled Library

CRISPR/Cas9 Synergistic Activation Mediator (SAM) is an engineered protein complex for the transcriptional activation of endogenous genes. It consists of three components:

1. A nucleolytically inactive Cas9-VP64 fusion

2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2 (in Addgene #61427)

3. The MS2-P65-HSF1 plasmid which expresses the activation helper protein

SAM can be combined with a human genome-wide library to activate all known coding isoforms from the RefSeq database (23,430 isoforms).

Library description: The SAM library consists of 3 unique sgRNAs targeting each human RefSeq coding isoform in the proximal promoter (> 90% of sgRNAs are targeted to the first 200bp upstream of the transcription start site of their target). The total library size is 70,290 guides. For SAM gain-of-function screening, this sgRNA library has to be combined with two additional SAM constructs – dCas9-VP64 and MS2-P65-HSF1.
These are provided along with the library.

Protocols & References

• Zhang Lab SAM Website

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. Naturedoi:10.1038/nature14136 (2014). Pubmed

Before ordering, please review the steps for using pooled lentiviral CRISPR libraries.

• Protocol for SAM pooled library ampilfication (306.2 KB)

• Protocol for screening SAM pooled library (171.3 KB)

• SAM target sequences (2.2 MB)

• SAM deep sequencing data (645.4 MB)

Delivery

Human SAM Library
(3 Plasmid System - SAM) (cat#1000000057)