SLC25A22 Knockout HCT116 Cell Line

The choice depends on whether you are studying SLC25A22 (glutamate carrier 1, GC1)'s role as a mitochondrial glutamate-H⁺ symporter or modeling early infantile epileptic encephalopathy type 3 (EIEE3). The Knockout line is the standard tool for asking whether GC1 is required for mitochondrial glutamate import — GC1 imports glutamate into mitochondria for catabolism, ammonia detoxification (urea cycle precursor supply), and oxidative phosphorylation substrate provision. Overexpression is useful for studying disease-associated mutations or for biochemical reconstitution. For mitochondrial glutamate metabolism research, the EDITGENE SLC25A22 Knockout in HCT 116 enables study of glutamate-dependent mitochondrial functions. SLC25A22 mutations cause EIEE3 (autosomal recessive, severe neonatal epilepsy with brain malformations) — disease variant rescue enables genotype-function correlation studies. Rescue with wild-type or transport-deficient GC1 is the standard specificity control.

Primary applications: • Mitochondrial glutamate transport: in vitro mitochondrial swelling assays or fluorescent glutamate flux measurements to quantify GC1 activity. • Glutamate-driven oxygen consumption: Seahorse-based glutamate-stimulated respiration as a readout of mitochondrial glutamate import capacity. • EIEE3 modeling: rescue with patient-derived SLC25A22 mutations for genotype-function correlation studies of this severe neonatal epilepsy. • Urea cycle and ammonia handling: studies of mitochondrial nitrogen metabolism in the absence of GC1. EDITGENE recommends this model for researchers investigating mitochondrial glutamate metabolism, early infantile epileptic encephalopathy mechanisms, and mitochondrial amino acid handling.

Yes. GC1 rescue experiments are well-established for mitochondrial glutamate biology: • Construct design: use a codon-modified SLC25A22 sequence with a small C-terminal tag (FLAG, HA). GC1 has the SLC25 canonical architecture. • Mitochondrial localization validation: confirm mitochondrial inner membrane localization. • EIEE3 disease mutation rescue: patient-derived SLC25A22 mutations (e.g., P206L, G236W, R155Q) enable disease genotype-function correlation. • Functional readout: rescue should restore mitochondrial glutamate import (in vitro swelling assays) and glutamate-supported respiration (Seahorse). HCT 116 transduces efficiently with lentivirus and supports stable rescue line generation.