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In vitro transcription (IVT) is the synthesis of RNA molecules. Creative Biolabs produces CAR mRNAs with our proprietary production platform, ensuring the quality and expression efficiency. This product is anti-CD117 scFv (XW-576)-28BBZ chimeric antigen receptor (CAR) mRNA which can be used for CAR expression in various immune cells.
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CAR Information
Target Species
Human
Clone
XW-576
Host
Human
CAR Construction
scFv-CD28-41BB-CD3ζ
CAR Generation
Third
CAR Signaling Description
CD28 CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatory signals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells (APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR+ T cells containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy. 41BB CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigen-nonspecific chimeric antigen receptor T-cells to significantly increases antitumor activity. CD3ζ CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ , ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta, which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.
CAR mRNA Properties
Cap
Cap 1
PolyA
Segmented PolyA
Product Type
CAR IVT mRNA
Concentration
Lot specific
Buffer
1 mM Sodium citrate (pH 6.4)
Storage
Aliquot and store at -80°C. Avoid freeze/thaw cycles.
Handling
Thaw and work with mRNA on ice. Upon first use, pulse spin before opening and aliquot into single use portions. DO NOT vortex. Use only certified RNase-free reagents and consumables with proper RNase-free technique. Use of barrier tips is recommended. Avoid freeze/thaw cycles. DO NOT mix with media containing serum unless first complexed with a stabilizing transfection reagent.
mRNA Quality Control
OD260/280
PASS
Agarose Gel Mobility
PASS
IVT Vector Sequencing
PASS
IVT Vector Enzyme Digestion
PASS
mRNA Transfection Efficiency
PASS
CAR Expression after mRNA Transfection
PASS
Target Information
Target
CD117
Gene Name
KIT
Full Name
KIT proto-oncogene, receptor tyrosine kinase
Introduction
This gene encodes a receptor tyrosine kinase. This gene was initially identified as a homolog of the feline sarcoma viral oncogene v-kit and is often referred to as proto-oncogene c-Kit. The canonical form of this glycosylated transmembrane protein has an N-terminal extracellular region with five immunoglobulin-like domains, a transmembrane region, and an intracellular tyrosine kinase domain at the C-terminus. Upon activation by its cytokine ligand, stem cell factor (SCF), this protein phosphorylates multiple intracellular proteins that play a role in in the proliferation, differentiation, migration and apoptosis of many cell types and thereby plays an important role in hematopoiesis, stem cell maintenance, gametogenesis, melanogenesis, and in mast cell development, migration and function. This protein can be a membrane-bound or soluble protein. Mutations in this gene are associated with gastrointestinal stromal tumors, mast cell disease, acute myelogenous leukemia, and piebaldism. Multiple transcript variants encoding different isoforms have been found for this gene.
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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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