TMEM65 Knockout HCT116-LUC Cell Line

The choice depends on whether you are studying TMEM65's role in mitochondrial cristae organization and respiratory chain maintenance, or modeling encephalomyopathy phenotypes. The Knockout line is appropriate for asking whether TMEM65 is required for mitochondrial function in colorectal cancer cells. Overexpression is useful for testing sufficiency or for rescue with disease-associated TMEM65 variants. For TMEM65 research, the EDITGENE Knockout in HCT116-LUC offers a unique advantage: the luciferase reporter enables in vivo bioluminescence imaging for xenograft studies, allowing researchers to assess how TMEM65 loss affects tumor growth and metabolism in animal models. Combined with mitochondrial function readouts (Seahorse, membrane potential), this provides a comprehensive in vitro/in vivo platform. Rescue with wild-type or disease-mutant TMEM65 enables structure-function and disease-modeling studies.

Primary applications: • Mitochondrial function: Seahorse OCR/ECAR measurements, mitochondrial membrane potential analysis (TMRM), and respiratory chain complex activity assays in the absence of TMEM65. • In vivo xenograft imaging: the luciferase reporter enables longitudinal bioluminescence imaging of tumor growth and metabolic phenotypes in mouse xenograft models. • Cristae organization: electron microscopy or imaging of mitochondrial morphology to assess TMEM65's reported role in cristae structure maintenance. • Cancer metabolism studies: integration of TMEM65 loss effects on mitochondrial metabolism with colorectal cancer biology readouts (proliferation, oxidative stress sensitivity). EDITGENE recommends this model for researchers investigating mitochondrial cristae biology, cancer metabolism, and in vivo tumor models requiring bioluminescence imaging.

Yes. TMEM65 rescue experiments require attention to mitochondrial inner membrane targeting: • Construct design: use a codon-modified TMEM65 sequence with a small C-terminal tag (FLAG, HA). TMEM65 contains an N-terminal mitochondrial targeting sequence — N-terminal tags must not disrupt mitochondrial import. • Localization validation: confirm mitochondrial localization of exogenous TMEM65 by immunofluorescence co-staining with TOM20 or by mitochondrial fractionation before functional assays. • Disease mutation rescue: TMEM65 mutations associated with mitochondrial encephalomyopathy can be introduced for genotype-function studies. • Functional readout: rescue should restore mitochondrial membrane potential (TMRM), respiratory capacity (Seahorse), and cristae morphology (electron microscopy). HCT116-LUC transduces efficiently with lentivirus; the luciferase reporter is maintained through standard rescue line generation, preserving in vivo imaging capability.