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Human Activity-Optimized CRISPR Knockout Library (2 sub-libraries in lentiCRISPRv1)
(Pooled Library #1000000067)

  • Purpose

    Note: This library has been replaced by an updated version (see Depositor Comments below).

    The Sabatini/Lander CRISPR pooled library is optimized for cleavage activity, in order to maximize the likelihood of gene knockout.

    The library is split into two sub-libraries, each containing 5 sgRNAs per gene.

  • Vector Backbone

    lentiCRISPR v1 (Plasmid #49535) - expresses Cas9

    Note: This plasmid has been discontinued, as an updated version is available. For confirmation of screen hits, Addgene encourages users to use the updated lentiCRISPR v2 (Plasmid #52961), which can be requested separately.

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000067 Two gRNA pooled sub-libraries in lentiCRISPR v1 1
Available to academic and nonprofits only.

Library Details

  • Species
    Human
  • Genes targeted
    18,166
  • gRNAs
    178,896
  • Controls
    1,000
  • 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.

Pooled library #1000000067 will be delivered as two tubes, each containing a pooled sub-library. Note that each subpool must be transformed separately.

  • Volume
    ∼15 µL
  • Concentration
    10 ng/µL

Resource Information

Depositor Comments

Note: This item has been discontinued. The Sabatini and Lander labs have released an updated version of the library with an additional tube of gRNAs here.

Library screening is performed by comparing sgRNA abundance of an initial cell population and a final cell population after cell growth, using massively parallel sequencing.

Figure 1. Diagram of Library Screening Process

When performing sgRNA validation in a Cas9-expressing cell line, the depositing laboratory recommends using plasmid pLenti-sgRNA (Addgene #71409).

Amplified grna population graph

Figure 2. sgRNA diversity after initial amplification

Initial grna population graph

Figure 3. sgRNA diversity after infection of a target cell population

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:

    Activity-Optimized Human CRISPR Pooled Library was a gift from David Sabatini and Eric Lander (Addgene #1000000067)
  • For your References section:

    Identification and characterization of essential genes in the human genome. Wang T, Birsoy K, Hughes NW, Krupczak KM, Post Y, Wei JJ, Lander ES, Sabatini DM. Science. 2015 Oct 15. pii: aac7041. PubMed 26472758