Gentarget is dedicated to support the immunotherapy research field, by providing the comprehensive product lines related to all aspects of immunotherapy. Note: All products are provided for Research Use only.
Our mission are: (1) provide the over-express lentivirus products for the immune response targets, the booster genes and the suppress genes; (2) provide the knockdown guaranteed shRNA lentivirus for specific immune targets; (3) provide the ready-to-use CRISPR knockout lentivirus sets for specific immune targets; (4) provide target specific CAR, TCR, TAC lentivirus as ready-to-use reagents for your target validation assays. (5) provide the premade stable cell lines for T cell killing efficacy assay and your specific cell line-based assays; (6) Upon service requests, provide the customized lentivirus for your desired CAR, TCR or any other desired gene editing lentivirus.
The lentivirus expresses an immune response gene. They can be used to generate, in your desired cell types, the target expression cell lines which can be used to verify your CAR-T cell’s recognition, binding and killing efficacy via that specific target / antigen.
Those ready-to-use lentiviruses are a pool of three target specific knockdown shRNAs with guaranteed knockdown a specific target.
Our CRISPR knock-OUT lentivirus:
Those ready-to-use lentiviruses are mix of Cas9 expression lentivirus with two target specific gRNA lentivirus. Simply add it into your cells, and select the cells with the desired genomic knock-out.
Our CRISPR gene Repair and knock-IN lentivirus:
The kit includes three parts: (1) the CAS9 expression Adenovirus, or the Integrase Deficient Lentivirus expressing Cas9 enzyme ; (2) the target specific gRNA lentivirus; (3) the repair donor template or the engineered knock-IN cassette using our preoperatory engineered IDLD (“Integration Deficient Lentivector Donor”) format. This system provides high-efficient targeted gene editing with low off-target, less or none side-effect insertional mutagenesis, which will greatly accelerate the gene editing / therapy fields.
An effective and simple immune cell killing assay that closely mimics the immunotherapy activity in vivo, can greatly facilitate the therapy development. We generated a wide range of Fluorescent labelled cancer cell lines. After co-cultured the TCR or CAR T cells (or any other engineered effector cells) with those fluorescent labelled target cancer cells, T cell killing potency (%) can be measured in a time-course via Flow Cytometer, Cello-meter, Guava Machines, WOLF cell sorter, xCELL-igence or other cell analyzer. T cell-mediated cytolysis resulted in the decreased fluorescent signal level as the function of cell dead in an effector-cell dose-dependent matter in real-time. It enables the fast screen immunotherapy constructs, cell ratio and other conditions to identify the optimal treatment protocols.
We also generated luciferase expression cancer cell lines which can also using for T cell killing assay via luciferase signal level change. And the fluorescent cell lines expressing a specific checkpoint gene, can be used to validate your CAR-T efficacy targeted at this checkpoint.
GenTarget’s premade lentivirus products are popular worldwide. With more than 20-year industry experience in lentivirus technology and novel cloning technology, and many our proprietary technologies, our experts can produce your desired lentivirus products in the fastest turnaround time and most competitive prices.
CRISPR is a powerful gene-editing technology that potentially allows the ability to target multiple genes in T cells to improve cancer immunotherapy. There are many CAR targets (see table down below) and many T cell therapy candidates with a focus on diseases that have a clearly established genetic cause. Gentarget can provides the customized immunotherapy reagents as service orders. Click here to Contact Us with you request.
What is Immunotherapy:
Immunotherapy is an emerging cancer treatment roughly including two aspects: (1) deliver the genetic modified T cells to patients to killing cancer cells; (2) promote the immune responses for cancer suppression.
The advantages of Immunotherapy are: (1) the targeted cancer cell treatment can be achieved because of immune system precision. (2) immunotherapy is live, continuous and dynamical treatment and memorized so it can be self-adjusted, long-term treatment corresponding to the cancer development; (3) T- cell therapy can kill solid tumors, which are difficult to treat with other immune therapies.
How immunotherapy works:
Human immune system is activated when any “foreign substances” are encountered. The immune response attacks and destroy anything containing the Foreign substances, like cancer cells. The principle of immunotherapy is:
activating, training or engineering patient’s immune cells in a lab and reintroducing them into the same patient, to find and attack cancer cells;
Stimulating, or boosting the natural defenses of your immune system so it works harder or smarter to find and attack cancer cells;
The main types of immunotherapy:
checkpoint inhibitors: Immune checkpoints are molecules on certain immune cells that need to be activated (or inactivated) to start an immune response. Cancer cells sometimes find ways to use these checkpoints to avoid being attacked by the immune system. Checkpoint inhibitors are the drug or antibody to block the signaling pathway that suppress the immune system, which enable immune effector cells to attack cancer cells more aggressively. It acts at the activation of immune response, not directly on cancer cells.
Genetically Engineered T Cells: This treatment is directly act on cancer cells including several methods: (1) express a CAR (Chimeric Antigen Receptor) which composed of an intracellular signaling domain and the linked extracellular domain derived from a tumor-specific antibody. The tumor specific receptor guilds the T cells to attack specific tumor type. The killing depends upon the desired signaling pathway. The ideal cases are the CAR target only present on tumor cells, not normal cells; (2) The T cell was modified at its endogenous TCR (T Cell Receptor ) gene to recognize a tumor specific antigen, which activate the native T cell killing signal pathway. The TCR method has less side-effects than CAR does who elevates a un-nature signal level resulted in the severe Cytokine release syndrome side-effect; (3) coupled with a tumor specific antigen binding domain (TAC: T-cell Antigen Coupler), (4)combined engineered T cells with the knockout of endogenous TCR with insertion of an anti-cancer factor so that the T cell can provided as the allogeneic adoptive anti-cancer treatment;
Cytokines: Cytokines encompass a variety of protein families including interleukins, interferons, growth factors, tumor necrosis factors, and colony stimulating factors. Cytokines act on every phase of the cancer immunity cycle, improving antigen priming, inflammation and hematopoiesis and other specific physiological response to external stimuli including cancer. cytokines can stimulate the immune cells to attack cancer.
Others: cancer immunotherapy have many other types of treatment, including: Monoclonal antibodies (raised from cancel specific antigen, attack a very specific part of cancer cell); Cancer vaccines; Oncolytic virus, enrich or enhance the NK cell, Macrophage cell and so on.
The tumor or cell type specific antigens are the targets for CAR-T cell recognition. The T cell surface expressed antigens (receptors) are targets for immune boosting or for anti-tumor boosting via target-over expression or suppression via knockdown or knockout.
1) PD-L1 /PDL-2 and PD-1 (Programmed death-1):
PD-L1 and PD-L2, are expressed on the surface of tumor cells. Both bind for PD-1 that expressed in T cell surface which inhibits T-cell activity. Blocking the PD-1 signal pathway can stop tumor’s immune escape, activate T cell immunoreactions to cancers.
Gentarget’s PD-L1 over-expression lentivirus: CAT#: LVP1077 and LVP1077-GP, express human PD-L1 gene in any desired cell types. It contains the N-term 6His tag which also allow to purified the mammalian cell expressed PD-L1 protein for antibody or vaccine development.
The PD-L1 signal cell line (CAT#: SC061-PDL1) can be used to validate your treatment targeted at this pathway.
shRNA knockdown human PD-1: CAT#:
CRISPR-PD1 knockout lentivirus: Simply add it into your T cell to achieve the knockout the PD1 gene. It mixed with the premade lentivirus of CAS9 and gRNA (PD1). CAT#:
2) LAG3 (Lymphocyte-activation gene 3):
LAG-3 expressed on the surface of both activated cytotoxic T cells and regulatory T cells. Cancer cells active LAG-3 leads to T-cell exhaustion and eroding the ability of T cells to kill tumor cells. Inactivation of LAG-3 allowed T cells to regain cytotoxic function.
CRISPR-LARG3 knockout lentivirus: Simply add it into your T cell to achieve the knockout the PD1 gene. It mixed with the premade lentivirus of CAS9 and gRNA (LARG3). CAT#:
3) TCR (T-Cell Receptor):
The T-cell receptor (TCR) is a protein complex found on the surface of T cells, or T lymphocytes that is responsible for recognizing major histocompatibility complex (MHC I) molecules. Many TCRs recognize the same antigen peptide and many antigen peptides are recognized by the same TCR. TCR consists mainly, of an alpha (α) chain and a beta (β) chain (encoded by TRA and TRB, respectively).
shRNA knockdown lentivirus: knockdown validated, mixed of 3 shRNA for human TCRα- and β-chain genes, carrying GFP-Puromycin dual selection: CAT#
CRISPR knockout lentivirus for TCRα- and β-chain genes: CAT#:
CRISPR for TCRα- and β-chain gene rearrangement lentivirus, CAT#:
CRISPR for targeting TCR / TAC lentivirus: upon request.
CTLA-4 /CD152 are expressed on the surface of T cells to maintain immune reaction balance. Tumor cells utilize the CTLA-4 pathway to suppress initiation of an immune response, resulting in decreased T-cell activation.
CTLA-4 over-expression lentivirus: CAT#: LVP877 (human) and LVP916 (mouse)
shRNA knockdown lentivirus: knockdown validated, mixed of 3 shRNA for human CTLA-4 gene carry GFP-Puromycin dual selection: CAT#
CRISPR-CTLA-4 knockout lentivirus: Simply add it into your T cell to achieve the knockout the CTLA-4 It mixed with the premade lentivirus of CAS9 and gRNA (CTLA-4). CAT#:
CD73 is a cell-surface enzyme on regulatory T cells. Cancer cells express CD73 to reduce antitumor immunity. Tumor-derived CD73 is a contributor to immune escape in cancer. Inhibition of CD73 activity can stimulate T-cell activity.
shRNA knockdown lentivirus: knockdown validated, mixed of 3 shRNA for human CD73 gene carry GFP-Puromycin dual selection: CAT#
CRISPR- CD73 knockout lentivirus: Simply add it into your T cell to achieve the knockout the CD73 gene. It mixed with the premade lentivirus of CAS9 and gRNA (CD73). CAT#:
6) IDO (Indoleamine-2,3-dioxygenase-1):
IDO is an intracellular enzyme for tryptophan metabolism which keeps T-cell immune responses under control. Tumor cells hijack this immunosuppressive process by up-regulate IDO activity. Blockade of IDO can restore cytotoxic T-cell function.
GITR is an activating receptor on the surface of T cells and other immune cells. Once exposure to tumor antigen activates a T cell, the number of GITR receptors on its surface increases. Activation of GITR signaling can help enhance immunity through the activation of cytotoxic T cells
GITR over-expression lentivirus: CAT#:
9) OX40 (CD134):
OX40 is an activating receptor expressed on the surface of activated cytotoxic T cells and regulatory T cells. On cytotoxic T cells, OX40 binds to its ligand (OX40L), resulting in stimulatory signals that promote T-cell reproduction, function, and survival, favorable to the antitumor immune response.
10) SLAMF7 (Signaling Lymphocytic Activation Molecule Family member 7):
SLAMF7 (CD319) is expressed on the surface of virtually all Natural Killer (NK) cells and subsets of other immune cells. SLAMF7 activates NK cells, the rapid responders of the immune system and the body’s first line of defense against cancer, which also initiate the development of long-term immunity by T Cell memory.
KIRs are expressed on the surface of Natural Killer (NK) cells and cytotoxic T cells. Normal cell expresses the ligand that inhibit KIRs that stop NK cells from killing normal cells. Tumor cells up-regulate the ligand for inhibitory KIRS in order to evade NK cell-mediated recognition and destruction. Blockade of inhibitory KIRs help restore NK cell-mediated immune activity.
CSF1R (CD115) is a cell-surface receptor expressed by macrophages and other cells. In some cancer, macrophages promote cancer survival. CSF1, the ligand for CSF1R, is a dominant regulator of macrophage differentiation and function. Cancer cell use upregulate CSF1 to target CSF1R on macrophages, supporting tumor growth. Blockade of CSF1R can deplete tumor-associated macrophages, thus improved T-cell responses.
shRNA knockdown lentivirus: knockdown validated, mixed of 3 shRNA for human CSF1R gene carry GFP-Puromycin dual selection: CAT#
CRISPR- CSF1R knockout lentivirus: Simply add it into your T cell to achieve the knockout the CSF1R gene. It mixed with the premade lentivirus of CAS9 and gRNA (CSF1R). CAT#:
Depend on cell type, CD 28 promote proliferation and function of T cell, also it serves anti-inflammatory roles in regulatory T cell (Treg). CD28 is the only B7 (B7.1: CD80, B7.2: CD86) receptor constitutively expressed on naive T cells.
“B-cell maturation antigen” or “tumor necrosis factor receptor superfamily member 17”:
BCMA is a cell surface receptor of the TNF receptor superfamily, preferentially expressed in mature B lymphocytes. TNFRSF17 is implicated in leukemia, lymphomas, and multiple myeloma and used as a target of CAR-T therapy.
TGFB1 is a secreted cytokine that performs many cellular functions. Regulatory T cells release TGF-β1 to inhibit the actions of other T cells and can inhibit the secretion and activity of many other cytokines. It also inhibits proliferation and stimulates apoptosis of B cells. TGF-β1 acts as a chemoattractant, directing an immune response to some pathogens.
shRNA knockdown lentivirus: knockdown validated, mixed of 3 shRNA for human TGFB1 gene carry GFP-Puromycin dual selection: CAT#
Lentivirus for over-expression CDs:
Gentarget also provide the premade lentivirus products express a cell surface CD antigen or receptor taht involves in intracellular signal-transduction pathways, active or suppress the immune response. See table below for those product name and Catalog number.
norvegicus CD59 molecule, complement regulatory protein
Lentivirus for over-expression Cytokines:
Cytokines encompass a variety of protein families including interleukins, interferons, growth factors, tumor necrosis factors, and colony stimulating factors. Cytokines act on every phase of the cancer immunity cycle, improving antigen priming, inflammation and hematopoiesis and other specific physiological response to external stimuli including cancer. Gentarget provides the ready to useover-expressing lentivirus for cDNAs including many cytokines. Please search the specific product on our home page.
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Moser, R.J., et.al; Curr. Gene Ther. 16, 207–219.
Stadtmauer et al., Science 367, 1001 (2020) 28 February 2020