The IMMT Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney cells, designed for targeted disruption of the IMMT gene. The polyclonal format provides a heterogeneous pool of edited alleles, enabling robust loss-of-function studies of IMMT, which encodes mitofilin, a core subunit of the mitochondrial contact site and cristae organizing system (MICOS). This product offers a reliable model for investigating IMMT-dependent mitochondrial biology in a widely used mammalian host.
HEK293T is an epithelial cell line originally generated by transformation of human embryonic kidney cells with sheared adenovirus type 5 DNA. It stably expresses the SV40 large T-antigen, conferring high transfection efficiency and supporting episomal replication of plasmids containing the SV40 origin. These features have established HEK293T as a standard platform for recombinant protein expression, viral vector production, and diverse cell biology applications, providing a well-characterized background for gene-editing studies.
IMMT is an essential structural protein that organizes mitochondrial cristae architecture via the MICOS complex. It interacts with SAMM50, CHCHD3, CHCHD6, OPA1, and mitofusins MFN1/MFN2 to maintain cristae junction integrity and inner membrane organization. IMMT expression is regulated by NRF1, PPARGC1A, and SP1, and its disruption leads to OPA1 processing defects, cristae destabilization, cytochrome c release, impaired ATP synthase activity, and enhanced sensitivity to intrinsic apoptosis. The mechanistic summary provided confirms that IMMT knockout disrupts the MICOS complex, causing mitochondrial inner membrane disorganization, reduced oxidative phosphorylation, and altered mitochondrial dynamics.
In HEK293T cells, IMMT knockout creates a tractable model system to dissect mitochondrial structure-function relationships. The parental line’s high transfection efficiency permits facile re-expression of wild-type or mutant IMMT for complementation and structure?Cfunction analyses. Moreover, the well-characterized proteomic landscape of HEK293T supports quantitative mass spectrometry and biochemical fractionation studies. This model thus enables precise correlation of molecular perturbations with changes in mitochondrial ultrastructure, metabolic flux, and apoptotic signaling, offering a versatile platform for mechanistic investigations in a human epithelial context.
These cells are suited for a range of applications including transmission electron microscopy of mitochondrial morphology, Seahorse metabolic flux analysis, mitochondrial membrane potential assays (JC-1, TMRM), and immunoblotting for MICOS components and apoptosis markers. Additional uses include co-immunoprecipitation of MICOS interactions, immunofluorescence localization, and functional apoptosis assays. The model is particularly relevant for research into mitochondrial disorders, neurodegeneration, cancer metabolism, and drug screening targeting mitochondrial integrity. For further information, please contact Ascent Research.
