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CRISPRi Guides for Enrichment-based Screening of Targets Library (GuEST-List)
(Pooled Library #248188, #248189)

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  • Purpose

    The CRISPRi Guides for Enrichment-based Screening of Targets Library (CRISPRi GuEST-List) enables genome-wide, single-cell pooled screens conducted with the Flex workflow from 10x Genomics. The gRNA protospacers are identical to those available in the Dolcetto A CRISPRi screening library from the Doench laboratory (Addgene #92385), containing a total of 57,050 constructs targeting over 18,000 human genes. The guide RNAs are encoded in a modified CROPseq vector backbone (Addgene #127458), whereby each guide RNA is associated with a unique pair of 25 base pair barcodes, which can be read out using a custom set of split probes compatible with the Flex kits. One library expresses a puromycin-resistance gene for selection, while the other library expresses both a puromycin resistance gene and eGFP.

  • Vector Backbone

    CROPseq-puro-v2 (Plasmid #127458) - does not encode a dCas9-CRISPRi effector

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 248188 CRISPRi GuEST-List v1† 1 $594 Add to Cart
Pooled Library 248189 CRISPRi GuEST-List GFP v1† 1 $594 Add to Cart
Available to Academic and Nonprofits Only

  • † A separate plasmid encoding a dCas9-CRISPRi effector is NOT included with this item. GuEST-List must be used with cell lines constitutively expressing a dCas9-CRISPRi effector, such as those established by lentiviral infection with Addgene #170068 or #188768.

Library Details

  • Species
    Human
  • Genes targeted
    18,901
  • gRNAs
    57,050
  • Controls
    500 unique non-targeting control gRNAs
  • 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.

  • Volume
    ∼20 µL
  • Concentration
    50 ng/µL

Resource Information

Depositor Comments

The GuEST-List plasmid design consists of a 5’ LTR, EF-1a promoter, puromycin resistance, P2A, plus or minus GFP, WPRE, and the 3’ LTR cassette made up of a hU6 promoter, sgRNA, barcode 1, and barcode 2. Genomic Integration results in the 3’ LTR cassette replacing the 5’ LTR and sgRNAs to be expressed from both of the 3’ LTR cassettes. Complementary Flex Probes (3’ to 5’) bind barcode 1 and 2 after puromycin selection.

Figure 1: GuEST-List Plasmid Design and Genomic Integration. Creative Commons Attribution License (Link opens in a new window).

Please visit https://doi.org/10.64898/2026.02.12.705613 (Link opens in a new window)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:

    Human CRISPRi GuEST-List Library was a gift from Rahul Satija (Addgene #248188 ; http://n2t.net/addgene:248188 ; RRID:Addgene_248188) Human CRISPRi GuEST-List GFP Library was a gift from Rahul Satija (Addgene #248189 ; http://n2t.net/addgene:248189 ; RRID:Addgene_248189)

  • For your References section:

    Genome-wide single-cell perturbation screens with VIPerturb-seq. Bradu A, Blair JD, Grabski IN, Mascio I, Lee J, McCormick C, Satija R. bioRxiv 2026.02.12.705613. doi: 10.64898/2026.02.12.705613
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