Description
Pre-made Lentivirus express the CAR construct of “3xFLAG-anti-CD19_CD20-ScFV (VL-VH)-CD8 hinge-4-1BB-CD3ζ”. It contains the GFP-Puromycin dual marker, allowing the selection of the transduced cells via antibiotic killing, or via GFP positive cell sorting.
This Bispecific Anti-CD19/CD20 CAR construct, contains the N-term 3xFLAG tagged “CD19-scFv-(G4S)x4 (clone FMC63)-CD20-scFv” recognition domain, linked to 4-1BB stimulatory domain, expressed under the enhance EF1a promoter. The lentivector’s core structure is represented by the scheme below.
CD19 is a marker of in most B cell leukemias and lymphomas but not in any normal tissue other than the B cell lineage. CD20 is another B cell surface protein involved in B cell activation and differentiation. Both CD19 and CD20 is used to diagnose cancers that arise from this type of cell, and used as the target for CD19/CD20-targeted CAR-T therapies (Reference). see details in Product Manual.
4-1BB (CD137) is a surface glycoprotein present on activated T Lymphocytes, CD4+ and CD8+ T cells. It binds to its ligand expressed on antigen-presenting cells (macrophages and activated B cells). It promotes T-cell antitumor activity. 4-1BB is one of the most widely studied co-stimulatory domain used in CAR design.
CD3ζ (T-cell receptor zeta) is expressed by T cells and NK cells. It together with T-cell receptor and CD3γ, δ, ε chain, forms the TCR-CD3 complex. CD3-zeta is the most commonly used activation component of CARs. It transmits an activation signal to the T cell after the antigen is bound. It can be coupled with additional co-stimulatory signaling for the complete activation.
Negative Controls: To validate the specificity of the targeted CAR-T killing, you can use the negative control CAR lentivirus: the CAR-ctr7 that has the same lentivector backbone but removed the Anti-CD-19/CD20 recognition.
Amount: 200 ul/vial at titer of 1×108 IFU/ml, concentrated lentivirus provided in PBS solution, premixed with 10x polybrene (60ug/ml), premade, ready to ship in dry-ice package.
CAT#: LVP1643
