Modular Pooled Knockin Libraries
(Pooled Libraries #207474, #207475, #207476, #207477, #207478, #207479, #207480, #207481)
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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.
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Vector Backbone
pUC19 - does not express Cas9
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Depositing Labs
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | ||
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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 |
Library Details
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SpeciesHuman
Pooled Library | Receptor | Genes Targeted | Number of Plasmids |
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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 |
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ControlsEach 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.
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Volume∼25 µL
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Concentration10 ng/µL
Resource Information
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Protocols
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Depositor Data
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Terms and Licenses
Academic/Nonprofit TermsIndustry Terms- Not Available to Industry
Trademarks- Zeocin® is an InvivoGen trademark.
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).
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.
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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.