NGFR(CD271,TNFRSF16) Chimeric Antigen Receptor (CAR): A Comprehensive Guide and Our Service & Product Introduction

NGFR (Nerve Growth Factor Receptor), also known as CD271 or TNFRSF16 (Tumor Necrosis Factor Receptor Superfamily Member 16), is a transmembrane glycoprotein belonging to the TNF receptor superfamily. It serves as a low-affinity receptor for neurotrophins and plays pivotal roles in neuronal development, cell survival, apoptosis, and tumor progression. As a professional provider of gene engineering tools, RGBiotech offers a comprehensive range of high-quality NGFR (CD271, TNFRSF16) CAR expression plasmid vectors and personalized custom construction services. We are dedicated to supporting researchers, biopharmaceutical enterprises in accelerating the development of NGFR (CD271, TNFRSF16)-targeted immunotherapies for various malignancies and diseases.
If you are interested in our NGFR (CD271, TNFRSF16) CAR expression plasmid vectors or custom construction services, or need technical advice for your NGFR (CD271, TNFRSF16) CAR research, please contact us at admin@rgbiotech.com. Our professional team will respond to your inquiries within 24 hours, providing personalized solutions to meet your research needs. We look forward to cooperating with you to advance NGFR (CD271, TNFRSF16) CAR technology and accelerate the development of innovative immunotherapies for NGFR-related solid tumors, cancer stem cells, and neurodegenerative diseases.

Our NGFR(CD271,TNFRSF16) CAR Expression Plasmid Vector Products and Custom Services

NGFR (CD271, TNFRSF16) CAR technology is a rapidly growing field in solid tumor immunotherapy and cancer stem cell targeting, with increasing demand for high-quality plasmid vectors and custom construction services. As a leading provider of gene engineering tools, RGBiotech offers a comprehensive range of NGFR (CD271, TNFRSF16) CAR expression plasmid vectors and professional custom construction services, tailored to meet the diverse needs of researchers and biopharmaceutical companies. Our products and services are designed to accelerate NGFR (CD271, TNFRSF16) CAR research and clinical translation, with strict quality control and full-cycle technical support.

Our NGFR (CD271, TNFRSF16) CAR expression plasmid vectors cover all generations (1st to 5th) and feature diverse designs to support various research applications.
1) 1st Generation: CD3ζ only (basic activation signal, low persistence, suitable for preliminary antigen recognition studies);
2) 2nd Generation: CD3ζ + one co-stimulatory domain (CD28 or 4-1BB, enhanced activation and proliferation, ideal for in vitro studies);
3) 3rd Generation: CD3ζ + two co-stimulatory domains (CD28+4-1BB/OX40, improved in vivo persistence and anti-tumor activity, suitable for in vivo models);
4) 4th Generation (ARM CAR): CD3ζ + co-stimulatory domain + cytokine (IL-7/IL-15/IL-21, enhanced anti-tumor immunity and tumor infiltration, suitable for solid tumor research);
5) 5th Generation: CD3ζ + co-stimulatory domain + inhibitory checkpoint antagonist (PD-1/CTLA-4 blockade, overcoming T cell exhaustion, ideal for long-term disease control).

Product Features

Product Applications

Custom NGFR (CD271, TNFRSF16) CAR Plasmid Vector Construction Services

Introduction of NGFR(CD271,TNFRSF16)

The NGFR gene (official symbol: NGFR; HGNC ID: 7809; Gene ID: 4804; also known as CD271, TNFRSF16, P75NTR, GP80-LNGFR, P75(NTR)) is located on human chromosome 17q21.33, spanning approximately 19.7 kb of genomic DNA. It is a protein-coding gene belonging to the TNF receptor superfamily, with a high degree of evolutionary conservation across mammalian species. The human NGFR gene has a reference sequence of NM_002507.3, with a gene length of 1311 bp, encoding a 75 kDa transmembrane glycoprotein composed of 427 amino acids. Multiple alternatively spliced transcript variants have been identified, producing distinct isoforms that differ in their cytoplasmic domains and exhibit diverse functional properties. The gene’s 5’-flanking region contains conserved transcription regulatory elements, including binding sites for transcription factors involved in neuronal development and tumor progression, leading to tissue-specific and pathological expression patterns.

NGFR (CD271, TNFRSF16) is a transmembrane glycoprotein with a structure specifically adapted for neurotrophin binding and signal transduction. Its structure consists of three core domains: an extracellular domain, a single transmembrane domain, and a cytoplasmic domain. The extracellular domain contains four 40-amino acid cysteine-rich repeats (each with 6 conserved cysteine residues) followed by a serine/threonine-rich region, which forms the neurotrophin-binding site-enabling low-affinity binding to nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5). The transmembrane domain anchors the protein to the cell membrane, while the cytoplasmic domain (155 amino acids in length) contains death domain motifs and binding sites for adaptor proteins (e.g., TRAF6, NRAGE), which mediate downstream signaling pathways related to cell survival, apoptosis, and differentiation. A soluble form of NGFR (sNGFR) is also produced by proteolytic cleavage of the extracellular domain, which regulates neurotrophin availability and signaling in the tumor microenvironment and inflammatory tissues.

NGFR (CD271, TNFRSF16) exhibits diverse functions depending on the cell type, ligand binding, and presence of co-receptors (e.g., TrkA). In normal physiology, it plays a critical role in neuronal development, axon guidance, neural crest cell migration, and tissue repair. It regulates the survival and apoptosis of neurons—promoting survival when bound to neurotrophins in the presence of TrkA, and inducing apoptosis in the absence of TrkA or when bound to pro-neurotrophins. Additionally, NGFR is involved in the regulation of mesenchymal stem cell (MSC) differentiation, hair follicle morphogenesis, skin development, and immune response modulation. In pathological conditions, abnormal NGFR overexpression promotes tumor cell proliferation, invasion, metastasis, and stemness, while also contributing to inflammation and tissue remodeling. It exerts these effects by activating downstream signaling pathways such as MAPK, PI3K-AKT, and NF-κB, which regulate cell cycle progression, epithelial-mesenchymal transition (EMT), and immune suppression.

In normal adult tissues, NGFR (CD271, TNFRSF16) expression is primarily restricted to cells of the nervous system, including neurons, glial cells, and neural crest-derived cells. It is also expressed at low levels in non-neuronal tissues, such as perivascular cells, dental pulp cells, lymphoid follicular dendritic cells, basal epithelium of oral mucosa, hair follicles, prostate basal cells, and myoepithelial cells of breast ducts. In contrast, NGFR is widely overexpressed in various malignant tumors, with high expression levels correlating with disease progression, metastasis, and poor prognosis. Notably, it is overexpressed in melanoma (particularly desmoplastic and neurotropic subtypes), neuroblastoma, pheochromocytomas, neurofibromas, colorectal cancer, pancreatic cancer, oral squamous cell carcinoma, and osteosarcoma. NGFR is also expressed on cancer stem cells (CSCs) in many of these tumors, making it an ideal target for targeted immunotherapy and CSC elimination.

NGFR (CD271, TNFRSF16) is closely associated with the development, progression, and prognosis of various tumors and non-malignant diseases, primarily due to its role in cell survival, apoptosis, and signaling.
1) Malignant Tumors: NGFR overexpression is a hallmark of multiple solid tumors, including melanoma (where it regulates tumor stemness and metastasis), neuroblastoma, neurofibromas, pheochromocytomas, colorectal cancer (where it regulates cancer stem cell dormancy and reawakening), pancreatic cancer, oral squamous cell carcinoma, and osteosarcoma. In melanoma, NGFR expression is required for tumor growth and metastasis, and its knockdown abolishes neural crest stem cell properties, reducing tumorigenicity and inducing apoptosis. High NGFR expression is also associated with resistance to targeted therapies (e.g., BRAF inhibitors) in melanoma, selecting for highly malignant metastatic cells.
2) Neurodegenerative Diseases: Abnormal NGFR expression is involved in neurodegenerative disorders such as Alzheimer’s disease, Parkinson’s disease, and amyotrophic lateral sclerosis (ALS), due to its role in neuronal apoptosis and neurotrophin signaling dysfunction.
3) Inflammatory and Autoimmune Diseases: NGFR dysregulation contributes to chronic inflammatory diseases (e.g., osteoarthritis, inflammatory bowel disease), as it limits inflammation and promotes tissue remodeling. Soluble NGFR (sNGFR) levels are elevated in inflammatory tissues, regulating neurotrophin availability and immune cell function.
4) Developmental Disorders: Mutations in the NGFR gene are associated with developmental disorders affecting the nervous system, including congenital insensitivity to pain with anhidrosis (CIPA), due to impaired neuronal survival and function.

Introduction of NGFR(CD271,TNFRSF16) Chimeric Antigen Receptor (CAR)

NGFR (CD271, TNFRSF16) Chimeric Antigen Receptor (CAR) is a genetically engineered receptor designed to redirect immune cells (e.g., T cells, NK cells, macrophages) to specifically recognize and eliminate NGFR (CD271, TNFRSF16)-expressing tumor cells and cancer stem cells. By fusing the extracellular NGFR-binding domain (e.g., anti-NGFR single-chain variable fragment (scFv)) with intracellular signaling domains, NGFR (CD271, TNFRSF16) CAR-equipped immune cells achieve MHC-independent recognition of NGFR-positive cells, overcoming the immunosuppressive tumor microenvironment and providing a promising strategy for treating NGFR-positive tumors and neurodegenerative diseases. Additionally, NGFR-derived hinges are used to facilitate efficient enrichment and detection of CAR-T cells in vitro and in vivo, enhancing the utility of NGFR CAR technology in preclinical and clinical research.

A typical NGFR (CD271, TNFRSF16) CAR consists of three core components, optimized for targeting NGFR-positive tumors and cancer stem cells.
1) Extracellular Antigen-Binding Domain: Usually a high-affinity anti-NGFR (CD271) scFv or NGFR ligand (e.g., NGF fragment), responsible for specific binding to the extracellular cysteine-rich repeats of NGFR on target cells. Fully human scFv domains are preferred to reduce immunogenicity and improve in vivo persistence. Monoclonal antibodies targeting NGFR (e.g., clone ME20, NTR/912) are commonly used as the basis for scFv development.
2) Hinge and Transmembrane Domain: Derived from CD28 or CD8α, or optimized NGFR-derived hinges (e.g., N3, N4), facilitating the stability of the CAR on the immune cell surface, enabling efficient magnetic cell sorting (MACS) enrichment and detection via flow cytometry, and reducing steric hindrance in the tumor microenvironment.
3) Intracellular Signaling Domain: Varies by CAR generation, including activation domains (CD3ζ) and co-stimulatory domains (CD28, 4-1BB, OX40, etc.), which regulate the activation, proliferation, persistence, and anti-tumor activity of CAR-engineered cells. Advanced designs may include cytokine fusions (IL-7, IL-15) or immune checkpoint antagonist domains to enhance efficacy in solid tumors and overcome T cell exhaustion.

Current Research Achievements of NGFR (CD271, TNFRSF16) CAR

Approved Drugs of NGFR (CD271, TNFRSF16) CAR

NGFR (CD271, TNFRSF16) CAR Research Hotspots

NGFR (CD271, TNFRSF16) CAR Research Difficulties & Challenges

Frequently Asked Questions (FAQs)

Q: What is the difference between different generations of NGFR (CD271, TNFRSF16) CARs, and how to choose the right one?
A: The main difference lies in the intracellular signaling domains. 1st generation CARs (CD3ζ only) are suitable for preliminary studies of NGFR recognition but have low in vivo persistence and are prone to exhaustion. 2nd (CD3ζ + single co-stimulatory domain) and 3rd (CD3ζ + two co-stimulatory domains) generations are ideal for preclinical studies requiring enhanced cell activation and persistence, especially for in vivo models targeting NGFR-positive tumors. 4th generation CARs (with cytokines) are suitable for solid tumor research to improve tumor infiltration and persistence, addressing the TME barrier. 5th generation CARs (with checkpoint antagonists) are designed to overcome T cell exhaustion, making them ideal for long-term disease control in advanced solid tumors. Choose based on your research goals (e.g., melanoma vs. neuroblastoma, in vitro vs. in vivo studies, cancer stem cell targeting) and cell type (T cells vs. NK cells vs. macrophages).

Q: Which vector backbone is suitable for NGFR (CD271, TNFRSF16) CAR-T/NK cell preparation, and why?
A: Lentiviral vectors are the most commonly used backbone for NGFR (CD271, TNFRSF16) CAR-T/NK cell preparation, as they can transduce both dividing and non-dividing cells, integrate into the host genome for stable long-term CAR expression, and have low immunogenicity-critical for avoiding human anti-mouse antibody (HAMA) responses. They are also compatible with NGFR-derived hinge designs for CAR cell enrichment. Retroviral vectors are suitable for dividing cells (e.g., activated T cells) and are cost-effective for large-scale production. AAV vectors are ideal for transient or tissue-specific NGFR CAR expression, with high safety and low integration risk. Non-viral plasmid vectors are suitable for in vitro studies or non-viral delivery systems (e.g., electroporation, LNP), reducing the risk of viral vector-related complications and lowering manufacturing costs—particularly for off-the-shelf therapies.

Q: How to ensure high NGFR (CD271, TNFRSF16) CAR expression efficiency in target cells?
A: Several factors affect expression efficiency: 1) Choose a suitable promoter (EF-1α for stable long-term expression, CMV for strong transient expression, or tumor-specific promoters for targeted expression); 2) Optimize the NGFR (CD271, TNFRSF16) CAR gene codons for the target cell type to enhance translation efficiency; 3) Use high-purity, low-endotoxin plasmids to avoid cytotoxicity that impairs cell viability and expression; 4) Select an appropriate delivery method; 5) Ensure the vector has a suitable linker (2A or IRES) for co-expression of CAR and markers, avoiding interference between components. For vectors with NGFR-derived hinges, ensure the hinge sequence is optimized for stability and enrichment efficiency.

Q: What are the key considerations for NGFR (CD271, TNFRSF16) CAR plasmid vector storage and handling?
A: To maintain plasmid integrity and activity: 1) Store plasmids at -20°C or -80°C, avoiding repeated freeze-thaw cycles (aliquot into single-use portions to prevent degradation); 2) Thaw plasmids on ice and centrifuge briefly before use to ensure uniform concentration; 3) Avoid exposure to high temperatures, UV light, and RNase/DNase contamination, which can damage plasmid DNA; 4) For lentiviral/retroviral backbones, store the packaged virus at -80°C with cryoprotectants to maintain transduction efficiency; 5) Follow sterile procedures when handling plasmids for cell culture applications to prevent contamination that could invalidate experiments; 6) Lyophilized plasmids can be stored at ambient temperature for three months, facilitating transportation and short-term storage.

Q: Can your custom service design NGFR (CD271, TNFRSF16) CAR vectors with specific modifications (e.g., NGFR-derived hinges, bispecific design, cytokine fusion)?
A: Yes. Our custom service supports various modifications, including integrating NGFR-derived hinges (e.g., N3, N4) to enable efficient CAR cell enrichment and detection, fusing cytokines (IL-7, IL-15, IL-21) to improve in vivo expansion and persistence, and designing bispecific CARs (e.g., NGFR/CD44v6) to mitigate tumor escape. Simply provide your modification requirements, and our team will design a tailored solution.

Q: How to verify the functionality of NGFR (CD271, TNFRSF16) CAR plasmid vectors after purchase?
A: We recommend the following verification steps: 1) Sequence verification to confirm the plasmid sequence is correct; 2) Transfect HEK293T cells with the plasmid, then detect NGFR (CD271, TNFRSF16) CAR expression via flow cytometry (using anti-CAR antibodies) or fluorescence microscopy (if fluorescent markers are included)-similar to the validation methods used for NGFR expression vectors; 3) Perform in vitro cytotoxicity assays using NGFR-expressing target cells (e.g., melanoma, neuroblastoma, colorectal cancer cell lines) to verify the killing ability of CAR-transduced T/NK cells; 4) Detect cytokine production (e.g., IFN-γ, IL-2) via ELISA to confirm CAR activation.

Q: What is the difference between NGFR (CD271, TNFRSF16) CAR-T and NGFR (CD271, TNFRSF16) CAR-NK cells, and which is better for my research?
A: NGFR (CD271, TNFRSF16) CAR-T cells have strong proliferation ability and long-term persistence, making them suitable for long-term disease control (e.g., advanced melanoma, colorectal cancer) but may have higher risk of CRS and difficulty infiltrating solid tumors. NGFR (CD271, TNFRSF16) CAR-NK cells have lower immunogenicity, no risk of graft-versus-host disease (GVHD), and rapid cytotoxicity, making them suitable for patients with compromised T cell function or acute advanced disease. They also have better penetration into solid tumors and lower CRS risk, with enhanced ability to target NGFR-positive cancer stem cells. However, CAR-NK cells have shorter in vivo persistence and may require repeated infusions. Choose based on your research focus (long-term control vs. rapid efficacy) and safety requirements (e.g., allogeneic vs. autologous therapy).

Q: How to reduce off-target effects of NGFR (CD271, TNFRSF16) CAR-engineered cells?
A: Off-target effects can be reduced by: 1) Using highly specific extracellular binding domains (e.g., fully human anti-NGFR scFv) that only recognize NGFR on tumor cells or cancer stem cells; 2) Using tumor-specific or cancer stem cell-specific promoters to restrict CAR expression to target cells, avoiding expression in normal NGFR-positive neuronal or perivascular cells; 3) Adding a suicide gene (e.g., iCasp9) to the vector to eliminate CAR-engineered cells if off-target toxicity occurs; 4) Designing bispecific CARs that target NGFR and a tumor-specific antigen, enhancing specificity for malignant cells; 5) Optimizing CAR affinity to avoid cross-reactivity with other TNF receptor superfamily members.

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