CNR1 Recombinant Monoclonal Antibody
Description
Structure and Function of CNR1
CNR1 is a 473-amino-acid protein with seven transmembrane domains, an extracellular N-terminal domain (amino acids 1–117), and an intracellular C-terminal domain (amino acids 401–473) . The extracellular region includes hydrophilic domains (e1, e2, e3), with e2 being essential for cannabinoid binding . CNR1 mediates effects of endocannabinoids (e.g., anandamide) and synthetic cannabinoids, influencing pain perception, appetite, and immune response .
Production of CNR1 Recombinant Monoclonal Antibodies
Recombinant monoclonal antibodies are engineered using recombinant DNA technology to ensure specificity and scalability:
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Immunization: Animals (e.g., mice) are immunized with recombinant CNR1 protein or synthetic peptides.
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Cloning: B-cell RNA is reverse-transcribed into cDNA, and antibody genes are cloned into vectors.
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Expression: Host cells (e.g., E. coli, mammalian cells) produce antibodies, purified via affinity chromatography .
Key Features:
Applications in Research and Diagnostics
CNR1 recombinant monoclonal antibodies enable precise detection and functional studies:
Table 1: Applications and Antibody Codes
Cancer and Metabolic Disorders
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Cancer: CNR1 overexpression correlates with malignancy in prostate cancer but better prognosis in hepatocellular carcinoma . Antibodies like AM251 inhibit tumor growth and induce apoptosis in melanoma and renal cancer models .
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Metabolism: CNR1 antagonists (e.g., Rimonabant) reduce obesity-related risks by targeting fat storage and insulin sensitivity . Antibodies help study CNR1’s role in glucose/lipid metabolism, as shown in HL-7702 cells .
Immune Regulation
CNR1 modulates immune responses by suppressing inflammation. Recombinant antibodies have been used to explore its role in autoimmune diseases and transplant rejection .
Clinical and Research Significance
CNR1 recombinant monoclonal antibodies bridge basic science and therapeutic development:
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Drug Discovery: Screening agonists/antagonists for pain management or obesity treatment .
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Biomarker Development: Identifying CNR1 expression patterns in diseases like inflammatory bowel disease .
Challenges and Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
CUSABIO has developed a rigorous process to produce the CNR1 recombinant monoclonal antibody with the highest precision. The process starts with the isolation of B cells from the spleen of an immunized animal using the recombinant human CNR1 protein as the immunogen. RNA is extracted from these B cells and converted into cDNA through reverse transcription. The gene encoding the CNR1 antibody is then amplified using a degenerate primer and inserted into a vector. This recombinant vector is introduced into host cells through transfection, facilitating the efficient expression of the CNR1 recombinant monoclonal antibodies. The expressed antibodies are harvested from the cell culture supernatant and purified using affinity chromatography. This antibody is specifically designed to recognize human CNR1 protein in ELISA.
Target Background
CNR1 is a G-protein coupled receptor for endogenous cannabinoids (eCBs), including N-arachidonoylethanolamide (also called anandamide or AEA) and 2-arachidonoylglycerol (2-AG), as well as phytocannabinoids, such as delta(9)-tetrahydrocannabinol (THC). It mediates a variety of cannabinoid-induced effects, including modulation of food intake, memory loss, gastrointestinal motility, catalepsy, ambulatory activity, anxiety, and chronic pain. Its signaling pathway typically involves a reduction in cyclic AMP. In the hypothalamus, CNR1 might have a dual effect on mitochondrial respiration depending on the agonist dose and potentially the cell type. It increases respiration at low doses, while decreasing respiration at high doses. At high doses, CNR1 signal transduction involves G-protein alpha-i protein activation, followed by inhibition of mitochondrial soluble adenylate cyclase, a decrease in cyclic AMP concentration, and inhibition of protein kinase A (PKA)-dependent phosphorylation of specific subunits of the mitochondrial electron transport system, including NDUFS2.
In the hypothalamus, CNR1 inhibits leptin-induced reactive oxygen species (ROS) formation and mediates cannabinoid-induced increases in SREBF1 and FASN gene expression. In response to cannabinoids, it drives the release of orexigenic beta-endorphin, but not melanocyte-stimulating hormone alpha/alpha-MSH, from hypothalamic POMC neurons, thereby promoting food intake. In the hippocampus, it regulates cellular respiration and energy production in response to cannabinoids. CNR1 is involved in cannabinoid-dependent depolarization-induced suppression of inhibition (DSI), a process in which depolarization of CA1 postsynaptic pyramidal neurons mobilizes eCBs, which retrogradely activate presynaptic CB1 receptors, transiently decreasing GABAergic inhibitory neurotransmission. It also reduces excitatory synaptic transmission.
In superior cervical ganglions and cerebral vascular smooth muscle cells, CNR1 inhibits voltage-gated Ca(2+) channels in both a constitutive and agonist-dependent manner. In cerebral vascular smooth muscle cells, cannabinoid-induced inhibition of voltage-gated Ca(2+) channels leads to vasodilation and decreased vascular tone. CNR1 induces leptin production in adipocytes and reduces LRP2-mediated leptin clearance in the kidney, contributing to hyperleptinemia. In adipose tissue, CNR1 signaling leads to increased expression of SREBF1, ACACA, and FASN genes. In the liver, activation by endocannabinoids leads to increased de novo lipogenesis and reduced fatty acid catabolism, associated with increased expression of SREBF1/SREBP-1, GCK, ACACA, ACACB, and FASN genes. CNR1 may also affect de novo cholesterol synthesis and HDL-cholesteryl ether uptake. Peripherally, it modulates energy metabolism. In high carbohydrate diet-induced obesity, CNR1 might decrease the expression of mitochondrial dihydrolipoyl dehydrogenase/DLD in striated muscles, as well as that of selected glucose/ pyruvate metabolic enzymes, thereby affecting energy expenditure through mitochondrial metabolism.
In response to cannabinoid anandamide, CNR1 elicits a proinflammatory response in macrophages, which involves NLRP3 inflammasome activation and IL1B and IL18 secretion. In macrophages infiltrating pancreatic islets, this process might participate in the progression of type-2 diabetes and associated loss of pancreatic beta-cells. CNR1 binds both 2-AG and anandamide; however, it only binds 2-AG with high affinity. Contrary to its effect on isoform 1, 2-AG acts as an inverse agonist on isoform 2 in assays measuring GTP binding to membranes. Similarly, it only binds 2-AG with high affinity, and 2-AG behaves as an inverse agonist on isoform 3 in assays measuring GTP binding to membranes.
- The cannabinoid receptor 1 (CNR1) was identified as a direct target gene of miR1273g3pand knockdown of CNR1 restored the phenotypes of LoVo cells transfected with miR1273g3p inhibitor. PMID: 29328379
- The Arg82 and Cys126 of CRIP1b are involved in the majority of hydrogen bond interactions with the CB1 receptor and are possible key residues required for interactions between the CB1 receptor and CRIP1b PMID: 28918320
- The CB1 pathway is involved in the pathogenesis of shoulder stiffness. It may be a promising target for the treatment of rotator cuff lesions with shoulder stiffness. PMID: 29108858
- CB1 receptors are among the most abundant G protein-coupled receptors in the brain that impact on several processes, including fear coping, anxiety, stress, learning, and memory. (Review) PMID: 29058339
- CB1 receptor (CB1b) is highly expressed in pancreatic beta-cells and hepatocytes but not in the brain. PMID: 27641999
- All of the epithelial layers in 94, 78, 96, 73 and 80% of pterygia cases, exhibited CB1, CB2, TRPV1, TRPV2 and TRPV3 cytoplasmic immunoreactivity, respectively. PMID: 28849159
- observed selective alterations of DNA methylation at the promoter of CNR1, the gene coding for the type-1 cannabinoid receptor, in schizophrenic patients with no changes in bipolar or major depressive disorder. In an animal model prefrontal cortex found a significant increase in CNR1 expression and a consistent reduction in DNA methylation at specific CpG sites of gene promoter. PMID: 28108228
- This study of single-nucleotide polymorphisms in the cannabinoid receptor gene CNR1 showed that C carriers at rs806374 may be at specific risk for increased odds of cannabis use during the transition out of high school (around age 18). PMID: 28930056
- results suggest that palmitoylation of the CB1 receptor finely tunes its interaction with G proteins and serves as a targeting signal for its functional regulation PMID: 28722168
- we have associated cannabinoid 1 (CB1) receptor genetic variation rs2180619 (AA, AG, GG), which is located in a potential CNR1 regulatory sequence, with performance in working memory. no differences were found among genotypes while performing each working memory (WM) task. However, the cost of the level of complexity in N-back paradigm was double for GG subjects than for AA subjects. PMID: 27108777
- two agonist-bound crystal structures of human CB1 in complex with a tetrahydrocannabinol (AM11542) and a hexahydrocannabinol (AM841) at 2.80 A and 2.95 A resolution, respectively PMID: 28678776
- that a genetically modulated balancing of signaling within the CB1-COX-2 pathway may reflect on more or less efficient patterns of prefrontal activity during working memory PMID: 27261878
- In cotransfected HEK-293 cells, SSTR5 and CB1R existed in a constitutive heteromeric complex under basal condition, which was disrupted upon agonist treatments. Furthermore, concurrent receptor activation led to preferential formation of SSTR5 homodimer and dissociation of CB1R homodimer. PMID: 27984180
- CB1R availability was inversely associated with BMI in homeostatic brain regions such as the hypothalamus and brainstem areas in both patients with food intake disorders(FID) and healthy subjects. However, in FID patients, CB1R availability was also negatively correlated with BMI throughout the mesolimbic reward system. PMID: 27404285
- hepatic expression of CB1 and CB2 plays an important role during the progression of fibrosis induced by chronic hepatitis B PMID: 28315398
- propose here a possible unifying theory of CB1 receptor functions. PMID: 28334603
- Patients with schizophrenia showed increased CB1A expression in cells of the innate immune system and simpler correlation network between cytokines and CBRs expression when compared with controls. PMID: 28011441
- our systematic review on potential associations of CNR1 polymorphisms with schizophrenia and its subphenotypes revealed conflicting data, with results tending towards a negative association. PMID: 28099629
- Our study disclosed an increased risk of CVS among individuals with AG and GG genotypes of CNR1 rs806380 ( P <0.01), whereas the CC genotype of CNR1 rs806368 was associated with a decreased risk of CVS ( P <0.05). In addition CT and CC genotypes of CNR1 rs806368 were associated with a family history of migraines. PMID: 28349993
- U46619 and ANG II may stimulate rapid endothelial release of endocannabinoids (mainly 2-arachidonoylglycerol), leading to CB1 receptor-dependent and/or CB1 receptor-independent vasorelaxation, which in the negative feedback mechanism reduces later agonist-induced vasoconstriction. PMID: 28356298
- Data suggest that signaling via endocannabinoids and CNR1/CNR2 plays central role in food intake; the endocannabinoid system regulates bidirectional communication (in the context of energy intake and storage) between the brain and peripheral tissues (adipose tissue, liver, gastrointestinal tract, endocrine pancreas, and skeletal muscle). [REVIEW] PMID: 28246151
- Selective estrogen receptor (ER) modulator (SERM) isomers exhibit distinct affinity and selectivity for cannabinoid subtype 1 and 2 receptors (CB1R and CB2Rs). PMID: 27936172
- We investigated the relationship between variation in the CNR1, CNR2, and FAAH genes and change in primary anxiety disorder severity. Five SNPs were nominally associated with a poorer treatment response (rs806365 [CNR1]; rs2501431 [CNR2]; rs2070956 [CNR2]; rs7769940 [CNR1]; rs2209172 [FAAH]) and one with a more favorable response (rs6928813 [CNR1]). PMID: 27346075
- we investigated whether the effect of the CB1 receptor coding CNR1 gene on migraine and its symptoms is conditional on life stress. We covered the CNR1 gene with 11 SNPs. None of the SNPs showed main genetic effects on possible migraine or its symptoms. 5 SNPs showed nominally significant interaction with recent negative life events on headache with nausea PMID: 27762084
- major allele homozygotes in rs806379 experienced significantly greater nicotine deprivation-induced reductions in cognitive control as indicated by enhanced alpha-1 resting EEG power. Analysis of the 'TAG' haplotype on CNR1 predicts reduced mRNA expression in postmortem analyses of the brain. The rs806379 findings suggest that CNR1 antagonism may mitigate the influence of nicotine deprivation on cognitive disruption. PMID: 27453054
- These findings suggest that Gi1 interacts only with active GPCRs and that the well known high speed of GPCR signal transduction does not require preassembly between G proteins and GPCRs. PMID: 28438833
- the presence and differential distribution of fatty acid amide hydrolase (FAAH) and monoglyceride lipase (MGLL) in relation to CB1 during the maturation of human oocytes, was investigated. PMID: 26948343
- CRIP1a can compete with beta-arrestins for interaction with C-terminal CB1R domains that could affect agonist-driven, beta-arrestin-mediated internalization of the CB1R. PMID: 27895162
- These findings indicate that D2 antagonism, like D2 agonists, change agonist-mediated CB1 coupling and signaling. PMID: 27053685
- there was a significant reduction in CB1 receptor gene expression levels in cancer tissue compared to normal surrounding mucosa of patients with CRC, confirming that in cancer the "protective" action of the CB1 receptor is lost. PMID: 28245562
- This study demonstrated that CB1 receptor up regulation in Human Stroke. PMID: 27425766
- We hypothesize that high expression levels of CNR1 provide tumor susceptibility to the antitumor effects of circulating endocannabinoids like anandamide, resulting in tumor involution of pediatric low-grade gliomas PMID: 27613640
- evaluation of CB1R expression status in biopsy specimens of esophageal squamous cell carcinoma using immunohistochemistry might be clinically useful for prediction of metastasis to lymph nodes and distant organs PMID: 27976824
- Data indicate that mobile tongue squamous cell carcinoma (SCC) patients presenting enhanced cannabinoid receptors CB1R, CB2R, or concomitant CB1R/CB2R expression showed significantly longer overall and disease-free survival times compared to those with low expression. PMID: 26459312
- This is the first demonstration that the fibroblasts of the muscular fasciae express CB1 and CB2. PMID: 27349320
- Epistatic interaction between the 1359 G/A polymorphism of the CNR1 gene and the DRD2 gene polymorphisms may be associated with cannabis addiction phenotype in a Turkish population. PMID: 26833047
- in the group of all HD patients, a significantly higher incidence of GA genotype and lower incidence in GG genotype in the polymorphism rs806381s were revealed versus the control group (p = 0.04). It seems that polymorphisms of the CNR1 gene are not associated with uremic pruritus. PMID: 27034934
- Results suggest that CNR1 SNP genotype is associated with increased total cholesterol and Myocardial Infarction. PMID: 25270105
- this study presents the 2.8 A crystal structure of human CB1 in complex with AM6538, a stabilizing antagonist, synthesized and characterized for this structural study. PMID: 27768894
- The cannabinoid 1 receptor polymorphism rs1049353, is involved in the processing of negative facial expressions with relation to anger in adolescence. PMID: 26527537
- Novel finding of the present study is the association of the mutant groups of CNR1 genotypes with lower levels of triglycerides and higher levels of HDL-cholesterol than the wild-type group. PMID: 25682784
- The present study reports an association of the G1359A and A1359A CNR1 genotypes with lower levels of triglycerides and higher levels of HDL-cholesterol in obese females. PMID: 25664923
- In the adult human ependyma, which usually shows central canal absence, CB1 may play a different role by modulating astrocyte functions PMID: 26634814
- Induction of apoptosis was mediated via CB1 as well as CB2, and expression of CB receptors was a prerequisite for therapy response in our models. PMID: 26775260
- CNR1 polymorphisms are not associated with obesity and metabolic disorders, including insulin resistance, in PCOS women. PMID: 24971918
- data support an association between endocannabinoid system activity (cb1) and alcoholism. PMID: 26253623
- Human lung-resident macrophages express CB1 and CB2 receptors whose activation inhibits the release of angiogenic and lymphangiogenic factors PMID: 26467187
- Studies indicate that crosstalk between cannabinoid receptor type 1 (CB1) and transient receptor potential vanilloid 1 (TRPV1) modulates pain and inflammation in arthritis. PMID: 26343051
- The minor C allele of rs2023239 showed an independent protective effect against lifetime Manic Depressive Disorder. PMID: 26331953
- CNR1 polymorphism and child physical abuse GxE interaction predicts adult PTSD threat symptoms. PMID: 26717543
