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Modular Pooled Knockin Libraries
(Pooled Libraries #207474, #207475, #207476, #207477, #207478, #207479, #207480, #207481)

  • Purpose

    Each Modular Pooled Knockin (ModPoKI) library is comprised of plasmids that include a T cell receptor (TCR) or chimeric antigen receptor (CAR) plus a gene of interest and homology arms for insertion into the TRAC locus of human T cells.

    Two sets of genes, transcription factors (TF) or cell surface receptors (SR), are available in combinations with the NY-ESO-1 TCR, CD19-BBz CAR, HA-GD2-28z CAR, or CD19-28z CAR. These libraries have been designed in a modular fashion, enabling a combinatorial set of 10,000 TF pairs (TFxTF) or screening both transcription factors and surface receptors (TF+SR) in a single screen.

    ModPoKI screening experiments with these libraries can be used to identify pathways to improve T cell immunotherapies and program other cellular functions.

  • Vector Backbone

    pUC19 - does not express Cas9

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 207474 plIG-106 NY-ESO-1 TCR SR Library 1 $380 Add to Cart
Pooled Library 207475 plIG-107 NY-ESO-1 TCR TF Library 1 $380 Add to Cart
Pooled Library 207476 plIG-109 CD19-BBz CAR SR Library 1 $380 Add to Cart
Pooled Library 207477 plIG-110 CD19-BBz CAR TF Library 1 $380 Add to Cart
Pooled Library 207478 plIG-123 HA-GD2-28z CAR TF Library 1 $380 Add to Cart
Pooled Library 207479 plIG-124 HA-GD2-28z CAR SR Library 1 $380 Add to Cart
Pooled Library 207480 plIG-128 HA-GD2-28z CAR TFxTF Library 1 $430 Add to Cart
Pooled Library 207481 plIG-132 CD19-28z CAR TF+SR Library 1 $380 Add to Cart
Available to Academic and Nonprofits Only

Library Details

  • Species
    Human
Pooled Library Receptor Genes Targeted Number of Plasmids
207474 NY-ESO-1 TCR Surface Receptors 129
207475 NY-ESO-1 TCR Transcription Factors 100
207476 CD19-BBz CAR Surface Receptors 129
207477 CD19-BBz CAR Transcription Factors 100
207478 HA-GD2-28z CAR Transcription Factors 100
207479 HA-GD2-28z CAR Surface Receptors 129
207480 HA-GD2-28z CAR Pairs of Transcription Factors ~10,000
207481 CD19-28z CAR Surface Receptors and Transcription Factors 229
  • Controls
    Each library also includes two control plasmids encoding GFP and RFP. The TFxTF Library #207480 contains four controls: GFP-GFP, RFP-RFP, GFP-RFP and RFP-GFP.

Library Shipping

This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

  • Volume
    ∼25 µL
  • Concentration
    10 ng/µL

Resource Information

Depositor Comments

Pooled non-viral knockin experiments can be performed by electroporating cells with the library along with Cas9 ribonucleoprotein with gRNA targeting the TRAC locus.

The plasmids consist of the respective TCR or CAR in combination with additional genes (library members), adapters, and homology arms. The surface receptor part of the libraries consists of synthetic chimeric receptors (also known as "switch receptors"), chemokine receptors, cytokine receptors, metabolic receptors, and stimulatory molecules. Barcoded adaptors facilitate pooled cloning and can also be quantified by simple amplicon sequencing.

More information is available in the publication or the preprint on bioRxiv (doi: 10.1101/2022.07.27.501186 (Link opens in a new window)). Please also refer to Roth et al. 2020 Cell (doi: 10.1016/j.cell.2020.03.039 (Link opens in a new window)) for more details, including bioinformatic analysis.

The NY-ESO-1 TCR sequence was provided by Antoni Ribas (University of California, Los Angeles); CD19-BBz CAR and CD19-28z CAR sequences were provided by Tobias Feuchtinger (Ludwig Maximilian University of Munich); HA-GD2-28z CAR sequence was provided by Crystal Mackall and Robbie Majzner (Stanford University).

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:

    Modular Pooled Knockin Library was a gift from Alexander Marson (Addgene # )
  • For your References section:

    Modular pooled discovery of synthetic knockin sequences to program durable cell therapies. Blaeschke F, Chen YY, Apathy R, Daniel B, Chen AY, Chen PA, Sandor K, Zhang W, Li Z, Mowery CT, Yamamoto TN, Nyberg WA, To A, Yu R, Bueno R, Kim MC, Schmidt R, Goodman DB, Feuchtinger T, Eyquem J, Jimmie Ye C, Carnevale J, Satpathy AT, Shifrut E, Roth TL, Marson A. Cell 2023 Sep 14;186(19):4216-4234.e33. doi: 10.1016/j.cell.2023.08.013. PMID: 37714135.