Trap1 Knockout TCMK1 Cell Line

The Trap1 Knockout TCMK1 Cell Line is a CRISPR/Cas9-edited loss-of-function cell line derived from the mouse kidney epithelial TCMK1 host. This product offers a defined genetic model to study the mitochondrial chaperone Trap1, encoded by the Trap1 gene. Through targeted gene disruption, the cell line enables dissection of Trap1-mediated mitochondrial biology without confounding off-target effects.

TCMK1 is an SV40-immortalized epithelial cell line isolated from C3H/He mouse kidney, retaining key renal tubular characteristics including barrier and transport functions. Its epithelial origin and immortalized state provide a robust and scalable platform for in vitro investigation of kidney cell physiology, oxidative stress responses, and metabolic regulation.

Trap1 is a mitochondrial matrix-resident HSP90 paralog that acts as a key regulator of mitochondrial integrity and cell survival. Under oxidative stress, Trap1 expression is induced by transcription factors HSF1 and PGC-1??. As a molecular chaperone, it interacts directly with cyclophilin D (CypD) to suppress mitochondrial permeability transition pore opening, preventing cytochrome c release and subsequent apoptosis. Trap1 also modulates electron transport chain complex activity and inhibits SRC kinase signaling. Co-chaperones such as HOP (STIP1) and CDC37 assist in client protein folding, while Trap1 itself contributes to the mitochondrial unfolded protein response. Consequently, Trap1 knockout disrupts these protective interactions, leading to mitochondrial dysfunction, elevated reactive oxygen species, and enhanced apoptotic priming.

In the kidney epithelial context, Trap1 is essential for mitigating damage from ischemia-reperfusion and oxidative insults, which are central to acute kidney injury and chronic kidney disease pathogenesis. The TCMK1-based Trap1 knockout model mimics the loss of mitochondrial proteostasis and heightened stress vulnerability observed in renal disorders. This cell line thus provides a physiologically relevant system to explore how mitochondrial chaperone networks influence renal epithelial cell fate, metabolic adaptation, and disease progression, including renal cell carcinoma where Trap1 is often dysregulated.

Researchers can employ this cell line for diverse functional studies. Standard assays include Western blotting and RT-qPCR to confirm Trap1 ablation and assess pathway components like Bcl-2 family proteins. Seahorse metabolic flux analysis quantifies changes in oxidative phosphorylation and glycolysis. Apoptosis assays (Annexin V), ROS detection, flow cytometry, co-immunoprecipitation, and immunofluorescence enable detailed characterization of mitochondrial function, protein interactions, and cell death signaling. Applications span mitochondrial biology, cancer drug resistance, metabolic disorders, and kidney disease modeling. For more information or to request custom cell line services, please contact Ascent Research.