The TRPV1 Knockout HEK293T Cell Line is a CRISPR/Cas9-edited human knockout cell line featuring targeted disruption of the TRPV1 gene in HEK293T cells. This stable loss-of-function model enables precise investigation of TRPV1-mediated signaling pathways by eliminating endogenous channel activity, allowing researchers to dissect thermal and chemical nociception mechanisms without interference. The cell line is well-suited for heterologous expression, pharmacological screening, and mechanistic studies.
HEK293T cells, derived from HEK293 cells, are a human embryonic kidney epithelial line stably expressing SV40 large T antigen, which enhances transient protein expression and plasmid replication. Their ease of transfection and robust growth make them a preferred host for ion channel and calcium signaling studies, offering a relevant cellular context for TRPV1 knockout analyses.
TRPV1 (transient receptor potential vanilloid 1) encodes a non-selective cation channel activated by capsaicin, heat (>43??C), and acidic pH. Its activity is modulated by upstream regulators such as NGF, bradykinin, PKA, PKC, and anandamide. Channel activation drives calcium influx, which triggers CGRP and substance P release, neuronal depolarization, and NF-??B activation. TRPV1 functionally interacts with PIP2, calmodulin, AKAP79/150, ??-arrestin, and Src kinase. Representative pathway components include TRPV1, PIP2, PKA, PKC, CaMKII, NF-??B, and MAP kinases, integrating pain and inflammatory signaling through calcium-dependent cascades.
In the HEK293T background, TRPV1 knockout abolishes capsaicin- and heat-induced calcium responses, eliminating downstream signaling events such as CGRP and substance P release, and NF-??B activation, thereby providing a clean null background. This model is ideal for reconstituting mutant TRPV1 variants, studying protein-protein interactions with factors like PIP2 and calmodulin, and testing pharmacological agents targeting TRPV1-mediated pathways. It directly addresses mechanisms of neuropathic pain, inflammatory pain, migraine, cancer pain, and thermal hyperalgesia in a tractable cellular system.
The knockout cell line supports diverse assays, including calcium flux measurement, patch clamp electrophysiology, capsaicin-induced calcium imaging, FLIPR assays, as well as biochemical techniques such as western blotting, RT-qPCR, and immunofluorescence to quantify pathway markers. Primary applications include pain research, high-throughput drug screening for TRPV1 antagonists, mechanism-of-action studies, and investigation of calcium signaling and neuroinflammation. For further information or to inquire about custom knockout cell line services, contact Ascent Research.
