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Mouse CRISPR Activation Pooled Library (Caprano P65-HSF)
(Pooled Libraries #1000000113, #92383, #92384)

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  • Purpose

    This mouse CRISPR activation library (Caprano P65-HSF) is in backbone pXPR_502 (P65-HSF). This Caprano library activates over 22,000 mouse genes and is used for genome-wide activation screening.

    Each gene activated by this library is targeted by 3–6 gRNAs. These gRNAs are split among the two pools: Set A (Cat #92383) contains gRNAs 1–3, Set B (Cat #92384) contains gRNAs 4–6. Each pool has a unique set of gRNAs, but both pools are designed to target the same genes. The slight discrepancy in number of genes targeted between Set A and Set B is because not all genes had 6 gRNAs.

    Depositor comments: The modified sgRNA tracr has two MS2 loops and two PP7 loops, although only the PP7 loops are used in this library. This library uses two vectors instead of 3, as the PP7-P65-HSF is on same vector as sgRNA itself. It was found that MS2-P65-HST (hygroR) vector gave poor titer. The 6 guides per gene were designed first based on location relative to TSS, then sequence preferences.

    Primers for the lentiGuide backbone can be used with this library in the Sequencing Protocol.

  • Vector Backbone

    pXPR_502 (Plasmid #96923) - uses P65-HSF to activate a gRNA-scaffold; does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000113 Two tubes, each with a half-library (Set A and B) in backbone XPR_502 (P65-HSF) †
  • #92383 (Set A)
  • #92384 (Set B)
 1 $540 Add to Cart
Pooled Library 92383 Set A, one tube, contains Mouse CRISPRa sgRNA library Caprano (P65-HSF) † 1 $325 Add to Cart
Pooled Library 92384 Set B, one tube, contains Mouse CRISPRa sgRNA library Caprano (P65-HSF) † 1 $275 Add to Cart
Available to academic and nonprofits only.

  • † A dCas9 activation plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with lenti dCAS-VP64_Blast (Addgene #61425).

Library Details

  • Species
    Mouse
  • Genes targeted
    22,774 (Set A), 22,658 (Set B)
  • gRNAs
    67,187 (Set A), 66,889 (Set B)
  • Controls
    500 unique non-targeting controls are in each of the two half-libraries (Set A and Set B)
  • Lentiviral Generation
    3rd

Library Shipping

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.

Pooled libraries #92383 and #92384 will be delivered as a single tube containing a pooled half-library with 3 gRNAs per gene.

Pooled library #1000000113 will be delivered as two tubes, each containing a pooled half-library with 3 gRNAs per gene (for a total of up to 6 unique gRNAs per gene).

  • Volume
    ∼20 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

Please visit https://www.biorxiv.org/content/early/2018/06/27/356626 for bioRxiv preprint.

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:

    Mouse Caprano CRISPR Activation Pooled Library (Set A and B) was a gift from David Root and John Doench (Addgene #1000000113)
    Mouse Caprano CRISPR Activation Pooled Library Set A was a gift from David Root and John Doench (Addgene #92383)
    Mouse Caprano CRISPR Activation Pooled Library Set B was a gift from David Root and John Doench (Addgene #92384)

  • For your References section:

    Optimized libraries for CRISPR-Cas9 genetic screens with multiple modalities. Sanson KR, Hanna RE, Hegde M, Donovan KF, Strand C, Sullender ME, Vaimberg EW, Goodale A, Root DE, Piccioni F, Doench JG. Nat Commun. 2018 Dec 21;9(1):5416. doi: 10.1038/s41467-018-07901-8. PubMed 30575746
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