Mouse CRISPR Activation Library (SAM 3-Plasmid System)
(Pooled Library #1000000075)

• Purpose:

  The mouse CRISPR/Cas9 Synergistic Activation Mediator (SAM) pooled library uses 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:

  lenti sgRNA(MS2)_puro optimized backbone (Plasmid #73797)

• Depositing Labs: Feng Zhang
• Publication: Joung et al Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016.

Library Details

• Species: Mouse
• Genes targeted: 23,439
• gRNAs: 69,225
• Controls: 491
• Lentiviral Generation: 3rd

Library Shipping

Each library is delivered in microcentrifuge tubes on blue ice. A tube's contents will not necessarily be frozen. For best results, minimize freeze/thaw cycles.

• Volume: ∼20 µL
• Concentration: 50 ng/µL


• You will also receive bacterial stabs of the lentiMPH v2 MS2-P65-HSF1 plasmid (Addgene #89308) and dCas9-VP64 (Addgene #61425). Please do NOT order these plasmids in addition to this library.

Resource Information

• Protocols:
  • For the most up-to-date and detailed instructions on using this library, please see Joung, et al. 2017 .
  • Library Amplification Protocol (PDF, 314 KB)
  • Screening Protocol (DOCX, 175 KB)
• Depositor Data:
  • Mouse SAM Target Sequences (CSV, 2.2 MB)
  • Mouse SAM Library Guide Sequences (CSV, 1.7 MB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

The SAM library consists of three 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).

For additional information, protocols, and an activator sgRNA design tool, visit the Zhang Lab website .

Figure 1: Diagram of SAM library constructs.