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PurposeModular CD28-CD3z CAR backbone, For Transient Expression or Lentiviral Production
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Depositing Lab
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Sequence Information
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Sequences (1) — Accept Affinity Reagent Sequence Policy
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Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 135991 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepSL
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Backbone manufacturerHeavily modified from LentiCRISPR v1 backbone
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Modifications to backboneUse BpiI single pot restriction ligation to swap the antigen binding domain of the CAR from CD19 to desired specificity
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Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameFMC63-28-3z-P2A-EGFP
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Alt nameCD19(FMC63)-CAR, CD28 costim., CD3z stim domains, P2A-GFP marker
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Alt nameContains: FMC63-scFV, Hinge domain, CD28-transmembrane domain, CD28-ITD, CD3z-ITD
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SpeciesH. sapiens (human)
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Entrez GeneCD28 (a.k.a. Tp44)
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Entrez GeneCD247 (a.k.a. CD3-ZETA, CD3H, CD3Q, CD3Z, CD3ZETA, IMD25, T3Z, TCRZ)
- Promoter EF1a-short
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BpiI (destroyed during cloning)
- 3′ cloning site BpiI (destroyed during cloning)
- 5′ sequencing primer CGGGTTTGCCGCCA
- 3′ sequencing primer caggagccTGCTCCTCTTACTcc (Common Sequencing Primers)
Resource Information
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Addgene Notes
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Article Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids 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:
pSLCAR-CD19-28z was a gift from Scott McComb (Addgene plasmid # 135991 ; http://n2t.net/addgene:135991 ; RRID:Addgene_135991) -
For your References section:
A High-Throughput Method for Characterizing Novel Chimeric Antigen Receptors in Jurkat Cells. Bloemberg D, Nguyen T, MacLean S, Zafer A, Gadoury C, Gurnani K, Chattopadhyay A, Ash J, Lippens J, Harcus D, Page M, Fortin A, Pon RA, Gilbert R, Marcil A, Weeratna RD, McComb S. Mol Ther Methods Clin Dev. 2020 Jan 31;16:238-254. doi: 10.1016/j.omtm.2020.01.012. eCollection 2020 Mar 13. 10.1016/j.omtm.2020.01.012 PubMed 32083149