TRPV1 Gene Editing    

TRPV1 channel, a member of the vanilloid TRP family, is a non-selective cation channel, which possesses significant permeability to ions including H⁺, Na⁺, Mg²⁺, and Ca²⁺. TRPV1 is a tetrameric ion channel, which is composed of intracellular amino, carboxy termini and putative six-transmembrane-domain with a loop helix domain localized between the 5th and 6th transmembrane domain. TRPV1 channel is a thermosensitive ion channel and possesses the ability to detect alterations in the environmental temperature (~42 °C). In addition, being heat sensitive, these channels are also activated by a broad array of noxious mechanical (pain, stretch), and chemical stimuli (arachidonic acid metabolites, acidic pH, bradykinin, toxins, such as resiniferatoxin and vanillotoxins) and blocked by agents including capsazepine, AMG 9810, SB366791, 5′-Iodoresiniferatoxin.

Figure 1. A schematic diagram showing the key structural features of TRPV1 receptors. (Aghazadeh Tabrizi M, et al. 2017)

Regulation and Distribution of TRPV1

TRPV1 is regulated by phosphorylation inducing an increased sensitivity to both thermal and chemical stimuli. In fact, TRPV1 receptors have multiple phosphorylation sites at various serine and threonine residues for protein kinase A (PKA), PKC, calcium-calmodulin kinase II (CaMKII) and c-Src kinase. Phosphorylation induced by PKA and PKC, via IP3 signaling and cyclic AMP signaling, respectively, produced different effects. In particular, PKA decreases TRPV1 desensitization capsaicin-mediated by phosphorylating Ser116, thereby allowing a higher sensitivity to the TRPV1 agonist. PKC phosphorylation instead, usually triggered by inflammatory agents, increases sensitization of this receptor to heat, agonists, or protons. In addition, PKC potentiates TRPV1 activity by shifting voltage-dependent activation to more negative potentials thus increasing the possibility of channel opening at normal membrane potentials.

According to its role in nociception, pain, and heat perception, TRPV1 expression has been originally detected in primary afferent nociceptors of the trigeminal ganglia (TG), dorsal root ganglia (DRGs), and vagal ganglia. Subsequent studies showed a much wider distribution in the central nervous system (CNS), for example, in dopaminergic neurons of the substantia nigra, hypothalamus, hippocampus, cerebellum, cortex, in dentate gyrus, and the nucleus accumbens. Moreover, TRPV1 is also expressed in nonneuronal cells, for example, epidermal keratinocytes, liver hepatocytes, urothelium, polymorphonuclear granulocytes, endothelial cells, pancreatic β-cells, mononuclear cells, arteriolar smooth muscle cells, preadipocytes, mesenteric arteries, and adipose tissue.

TRPV1 and Cancer Therapy

Functional expression of TRPV1 was demonstrated in several tumor types including human papillary thyroid carcinoma BCPAP cells, human breast cancer cell lines (MCF-7 and BT-20), prostate cancer (LNCaP and PC-3), urothelial cancer cells, and glioma. Several studies addressed the TRPV1 activation in anti-cancer therapy by harnessing the Ca²⁺ signaling. Activation of TRPV1 by capsaicin can significantly reduce proliferation and induce apoptosis of aggressive triple-negative breast cancer cell line (SUM149PT). Capsaicin itself may not only play a role through TRPV1-dependent calcium signaling pathway. For example, Hawng et al. showed that the co-carcinogenic action of capsaicin in TPA-promoted skin carcinogenesis is related to the activation of the EGFR/Akt pathway not Ca²⁺ signaling via TRPV1. Similarly, Bao et al. demonstrated that in the osteosarcoma MG63 cells capsaicin treatment increases phosphorylation of p53 and AMPK in the TRPV1-independent manner.

Several studies have shown that administration of chemotherapy along with TRPV1 activator—capsaicin, can produce synergistic effect, leading to increased apoptosis and suppression of tumor cell migration. Studies by Deveci et al. showed that the activation and apoptosis levels of TRPV1 in MCF-7 human breast cancer cell line were significantly higher than those in cells treated with 5-fluorouracil alone. Similarly, Nur et al. revealed that the apoptotic effect of cisplatin was improved by activation of TRPV1 in the MCF-7 cell line. Moreover, alteration of TRPV1 activity by capsaicin was shown to significantly reduce the migration and invasion of human papillary thyroid carcinoma BCPAP cells.

TRPV1 Gene Editing Services

CRISPR/Cas9 Platform^CB, one of the leading biotechnological companies specializing in gene editing, is dedicated to offering comprehensive CRISPR/Cas9 gene-editing services to a wide range of genomics researchers. Based on our platform, we can help you effectively TRPV1 gene deleted, inserted or point mutated in cells or animals by CRISPR/Cas9 technology.

• TRPV1 Gene Knockout: We offer TRPV1 gene knockout cell line and knockout animal model generation service with high quality. Typically, we develop CRISPR-mediated gene editing cell lines including HEK239T, Hela, HepG2, U87, but we can use other cell lines according to your requirements. Our one-stop KO animal model generation service covers from sgRNA design and construction, pronuclear microinjection to Founders genotyping and breeding.
• TRPV1 Gene Knockin: CRISPR/Cas9 Platform^CB provides the one-stop TRPV1 knock-in cell line and knockout animal model generation services, including point mutation and gene insertion. Our expert staff has succeeded in dozens of TRPV1 knock-in cell line generation projects, including stem cells, tumor cells and even difficult-to-handle cells. We also have extensive experience in incorporating CRISPR/Cas9 technology into animal models, which have been fully recognized by our clients.

If you have any questions, please feel free to contact us.

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