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htCRISPRi-Puro Library
(Pooled Library #254479, #254480)

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  • Purpose

    This library is a compact CRISPRi library (two sgRNAs per gene) targeting all genes expressed in human T cells. It was selecting from hCRISRPi-v2 and the Dolcetto libraries. The library was also designed for compatibility with 10x FLEX workflow for genome-wide Perturb-seq as part of the CZI Billion Cells Project.

  • Vector Backbone

    lentiGuide-Puro-PJY390 (Plasmid #251159) - does not express Cas9

    lentiGuide-Puro-P2A-EGFP-PJY392 (Plasmid #251161) - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 254479 htCRISPRi-Puro† 1 $511 Add to Cart
Pooled Library 254480 htCRISPRi-Puro-GFP† 1 $511 Add to Cart
Available to Academic and Nonprofits Only

  • † A Cas9 plasmid is NOT included with this item and will have to be ordered separately. It can be used in conjunction with pZR316_Lenti-Ef1a-ZIM3-dCas9-2A-BsdR (Addgene #252529), pHR-SFFV-dCas9-BFP-KRAB (Addgene #46911), or pHR-SFFV-KRAB-dCas9-P2A-mCherry (Addgene #60954).

Library Details

  • Species
    Human
  • Genes targeted
    12,779
  • gRNAs
    26,504
  • Nontargeting controls
    992
  • 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

Deconvolution algorithms for analyzing NGS data: Extract guide sequences from sequencing reads by running PoolQ with the search prefix “CACCG”, and count reads by alignment to a reference file of all possible guides present in the library.

It’s compatible with any existing pipelines such as MAGeCK (Link opens in a new window).

Please visit https://doi.org/10.64898/2025.12.23.696273 (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:

    htCRISPRi-Puro library was a gift from Alexander Marson (Addgene #254479 ; http://n2t.net/addgene:254479 ; RRID:Addgene_254479)
    htCRISPRi-Puro-GFP library was a gift from Alexander Marson (Addgene #254480 ; http://n2t.net/addgene:254480 ; RRID:Addgene_254480)

  • For your References section:

    Genome-scale perturb-seq in primary human CD4+ T cells maps context-specific regulators of T cell programs and human immune traits. Zhu R, Dann E, Yan J, Retana JR, Goto R, Guitche RC, Petersen LK, Ota M, Pritchard JK, Marson A. bioRxiv. 2025. doi: 10.64898/2025.12.23.696273.
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