Description
Pre-made Lentivirus express the CAR construct of “3xFLAG-anti-TGFβ-ScFV (VL-VH)-CD8 hinge-4-1BB-CD3ζ”. It contains the Puromycin marker allowing the selection of the transduced cells via antibiotic killing.
This CAR construct, contains the N-terminal 3xFLAG tagged “TGFβ-scFv [VH-(G4S)x4-VL]” 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.
TGF-β is often associated with immunosuppressive effects in the tumor microenvironment. High levels of TGF-β can contribute to immune evasion by inhibiting the activity of cytotoxic T cells and promoting the generation of regulatory T cells (Tregs). To counteract the immunosuppressive effects of TGF-β in the tumor microenvironment, the CAR-T cells are developed to resist TGF-β-mediated suppression. This CAR construct assembled the coding sequence from some identified anti-human TGFβ antibodies (see 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-ctr1 that has the same lentivector backbone but removed the recognition domain.
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#: LVP1672
