MECR Knockout HEK293T Cell Line

The choice depends on whether you are studying MECR (mitochondrial trans-2-enoyl-CoA reductase)'s role as the terminal enzyme of mitochondrial fatty acid synthesis II (mtFASII) or modeling MEPAN (MECR-associated neurodegeneration) syndrome. The Knockout line is the standard tool for asking whether MECR is required for these processes — MECR catalyzes the NADPH-dependent reduction of mitochondrial trans-2-enoyl-ACP intermediates during de novo fatty acid synthesis within mitochondria, distinct from the cytosolic FAS pathway. mtFASII generates octanoyl-ACP, the substrate for lipoic acid synthesis (essential for PDC, KGDH, BCKDH function). Overexpression is useful for studying MECR in heterologous expression contexts. For mitochondrial metabolism research, the EDITGENE MECR Knockout in HEK293T is highly informative — HEK293T's very high transfection efficiency supports systematic structure-function studies. MECR biallelic loss-of-function mutations cause MEPAN (autosomal recessive childhood-onset dystonia with optic atrophy and basal ganglia abnormalities, with bilateral basal ganglia degeneration on MRI) — disease variant rescue enables genotype-function studies. Rescue with wild-type or catalytically-dead MECR enables comprehensive structure-function studies. The knockout is valuable for studying lipoic acid biosynthesis, PDC/KGDH lipoylation, and emerging mtFASII-related therapeutic strategies.

Primary applications: • Lipoic acid biosynthesis: lipoylation status of PDC (PDHA1), KGDH (DLST), BCKDH (DBT), and GCS (GCSH) E2/H components by anti-lipoyl Western blot — mtFASII loss reduces lipoylation. • Mitochondrial bioenergetics: Seahorse OCR/ECAR analysis given mtFASII's contribution to mitochondrial complex function via lipoylation. • MEPAN modeling: rescue with patient-derived MECR mutations for genotype-function correlation studies of MECR-associated neurodegeneration. • mtFASII pathway analysis: ACP (mt-ACP) acyl chain composition by mass spectrometry to characterize mtFASII flux in the absence of MECR. EDITGENE recommends this model for researchers investigating mitochondrial fatty acid synthesis, lipoic acid biology, MEPAN disease mechanisms, and mtFASII-related metabolic disease.

Yes. MECR rescue experiments require attention to mitochondrial targeting: • Construct design: use a codon-modified MECR sequence with a small C-terminal tag (FLAG, HA). MECR has N-terminal mitochondrial targeting sequence cleaved upon import — N-terminal tags must not disrupt processing. • Mitochondrial localization validation: confirm mitochondrial matrix localization by appropriate compartment markers. • Catalytically-dead rescue: NADPH-binding or substrate-binding pocket mutations abolish enoyl-CoA reductase activity and serve as the standard specificity control. • MEPAN mutation rescue: patient-derived MECR mutations enable disease genotype-function studies. • Functional readout: rescue should restore PDC/KGDH/BCKDH lipoylation status (anti-lipoyl Western blot) and mitochondrial respiration as functional outputs of restored mtFASII. HEK293T transduces with very high efficiency and supports systematic structure-function rescue experiments.