Skip to main content
Addgene

Human CRISPR Activation Library (SAM - 3 Plasmid System)
(Pooled Libraries #1000000057, #1000000074)

  • Purpose

    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:

    1. A nucleolytically inactive Cas9-VP64 fusion (Addgene plasmid # 61425)
    2. A gRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2 (present in the libraries)
    3. 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

  • Vector Backbone

    For the zeocin resistant library, the backbone is lenti sgRNA(MS2)_zeo backbone (Plasmid #61427).

    For the puromycin resistant library, the backbone is lenti sgRNA(MS2)_puro backbone (Plasmid #73795).

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000057

Zeocin Resistant gRNA Pooled Library + 2 Helper Plasmids

1 $540 Add to Cart
Pooled Library 1000000074

Puromycin Resistant gRNA Pooled Library + 2 Helper Plasmids

1 $540 Add to Cart
Available to Academic and Nonprofits Only
  • A Cas9 plasmid IS included with this item.

Library Details

  • Species
    Human
  • Genes targeted
    23,430
  • gRNAs
    70,290
  • Lentiviral Generation
    3rd

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

  • Volume
    ∼20 µL
  • Concentration
    50 ng/µL

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.

Resource Information

Depositor Comments

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 200 bp 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.

Diagram of SAM library constructs. Guide RNA lentiviral expression vectors contain Psi+, RRE, cPPT, U6 promoter, sgRNA, MS2 stem loops, EF1 alpha promoter, puromycin or zeocin resistance, and WPRE. dCas0-VP64 lentiviral expression vectors contain Psi+, RRE, cPPT, EF1 alpha promoter, dCas9 with D10A and N863A mutations, NLS, VP64, P2A, blasticidin resistance, and WPRE. MS2-p65-HSF1 lentiviral expression vectors contain Psi+, RRE, cPPT, EF1 alpha promoter, MS2, NLS, P65, HSF1, P2A, hygromycin resistance, and WPRE.
Figure 1: Diagram of SAM library constructs
How to cite this pooled library ( Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    Human CRISPR Zeocin-Resistant 3-plasmid Activation Pooled Library (SAM) was a gift from Feng Zhang (Addgene #1000000057)
    Human CRISPR Puromycin-Resistant 3-plasmid Activation Pooled Library (SAM) was a gift from Feng Zhang (Addgene #1000000074)
  • 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 10 PubMed 25494202