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Anti-CAIX (Girentuximab)-FcRɣ+PI3K CAR-MA (CARMA-W0180)

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

The vector of anti-CAIX chimeric antigen receptor (CAR) is constructed for the engineering of macrophage cells to target human CAIX and induce target cell phagocytosis. The macrophage cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CAIX antibody (Girentuximab) linked to FcRɣ+PI3K signaling domains. The vector product was designed for the treatment of Metastatic clear cell renal cell carcinoma (ccRCC).

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Details

  • Target
  • CAIX
  • Cell Type
  • Macrophage
  • Targeting Diseases
  • Metastatic clear cell renal cell carcinoma (ccRCC)
  • Vector Name
  • pCDCAR1
  • Vector length
  • ~8kb
  • Vector Type
  • Lentiviral vector
  • Size
  • 10 µg
  • Storage
  • Store at -20°C for long term.

CAR Components

  • Promoter
  • EF1a
  • scFv
  • Girentuximab
  • Hinge
  • CD8
  • TM
  • CD8
  • ICD
  • FcRɣ+PI3K
  • Marker
  • GFP or N/A
  • Receptor Construction
  • scFv-hinge(CD8)-TM(CD8)-ICD(FcRɣ+PI3K)
  • Discription of Signaling Cassetes
  • FcRɣ
    FcRɣ is used as the CAR intracellular signaling domain. As a phagocytosis receptor, FcRɣ has cytosolic Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) that are phosphorylated by Src family kinases. Based on this mechanism, FcRɣ signaling can induce macrophage phagocytosis which has been demonstrated in the published article.
    Fc gamma receptors belong to the group of non-catalytic tyrosine-phosphorylated receptors which share a similar signalling pathway involving phosphorylation of tyrosine residues. The receptors generate signals within their cells through an important activation motif known as an immunoreceptor tyrosine-based activation motif (ITAM). An ITAM is present in the intracellular tail of FcγRIIA, and its phosphorylation induces phagocytosis in macrophages. FcγRI and FcγRIIIA do not have an ITAM but can transmit an activating signal to their phagocytes by interacting with another protein that does. This adaptor protein is called the Fcγ subunit and, like FcγRIIA, contains the two YXXL sequences that are characteristic of an ITAM.

    PI3K
    Previous work demonstrated that PI3K signaling is important for internalization of large targets. To increase PI3K recruitment to the CAR-P, we fused the portion of the CD19 cytoplasmic domain (amino acids 500 to 534) that recruits the p85 subunit of PI3K to the CAR. A CAR-P containing the p85 recruitment motif alone (CAR-P-PI3K) was able to induce some whole cell engulfment. Assembling an array of motifs designed to recruit distinct phagocytic effectors can increase CAR-P activity.

Target

  • Target
  • CAIX
  • Target Species
  • Human
  • Clone
  • Girentuximab
  • Host
  • Mouse
  • Gene Name
  • Carbonic anhydrase 9
  • Introduction
  • Carbonic anhydrases (CAs) are a large family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide. They participate in a variety of biological processes, including respiration, calcification, acid-base balance, bone resorption, and the formation of aqueous humor, cerebrospinal fluid, saliva, and gastric acid. They show extensive diversity in tissue distribution and in their subcellular localization. CA IX is a transmembrane protein and is one of only two tumor-associated carbonic anhydrase isoenzymes known. It is expressed in all clear-cell renal cell carcinoma, but is not detected in normal kidney or most other normal tissues. It may be involved in cell proliferation and transformation. This gene was mapped to 17q21.2 by fluorescence in situ hybridization, however, radiation hybrid mapping localized it to 9p13-p12.
  • Alternative Names
  • MN; CAIX; CA9; Carbonic anhydrase 9

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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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