MMACHC Knockout Cell Line (HEK293)

The choice depends on whether you are studying MMACHC's role as the cytosolic processor of dietary vitamin B12 (cobalamin) or modeling cblC (combined methylmalonic acidemia and homocystinuria) disease. The Knockout line is the standard tool for asking whether MMACHC is required for B12 processing — MMACHC catalyzes the dealkylation (removal of upper axial cyanide/methyl ligand) of dietary cobalamin precursors, generating cob(II)alamin that subsequently feeds into cytoplasmic methylcobalamin (for methionine synthase) or mitochondrial adenosylcobalamin (for MMUT) cofactor pathways. Overexpression is useful for studying MMACHC in heterologous expression contexts. For B12 metabolism research, the EDITGENE MMACHC Knockout in HEK293 is highly relevant — MMACHC mutations cause cblC disease, the most common inherited disorder of cobalamin metabolism, characterized by combined methylmalonic aciduria and homocystinuria due to defective AdoCbl and MeCbl production. Rescue with wild-type or patient-derived mutant MMACHC (e.g., c.271dupA, c.394C>T, c.481C>T) enables disease genotype-function studies. The knockout is valuable for studying B12 cofactor processing, cblC therapy (hydroxocobalamin, betaine), and B12 supplementation pharmacology. Complements the parallel MMUT Knockout (also available) for upstream-downstream B12 pathway dissection.

Primary applications: • Combined MMA and homocysteinemia modeling: cellular methylmalonic acid AND homocysteine accumulation (distinguishing cblC from isolated MMA) by LC-MS. • B12 cofactor synthesis: AdoCbl and MeCbl quantification downstream of MMACHC processing. • cblC disease modeling: rescue with patient-derived MMACHC mutations (c.271dupA most common, c.394C>T, c.481C>T) for genotype-function studies. • Hydroxocobalamin/betaine therapy: response to cblC treatment regimens in cellular models. EDITGENE recommends this model for researchers investigating cblC disease mechanisms, vitamin B12 processing, and B12-related metabolic disease therapeutics. Complements the parallel MMUT Knockout (also available) for upstream-downstream pathway studies.

Yes. MMACHC rescue experiments are well-established for cblC disease research: • Construct design: use a codon-modified MMACHC sequence with a small C-terminal tag (FLAG, HA). MMACHC has B12-binding pocket and reductase-like architecture — preserve catalytic residues. • Catalytically-dead rescue: B12-binding residue mutations or reductase active site mutations abolish cobalamin processing activity. • cblC mutation rescue: patient-derived MMACHC mutations enable disease genotype-function studies — the c.271dupA mutation is the most common cblC allele globally. • Functional readout: rescue should restore B12 processing (AdoCbl and MeCbl generation) and reduce methylmalonic acid and homocysteine accumulation. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.