Cobalamin-C defect is the most common inborn error of intracellular cobalamin metabolism. The nonsense mutation, c.394C>T (R132*), resulting in truncated protein, was common in North Indian cohort. Based on the location of mutation, it can be targeted for readthrough drug therapeutics for restoring the MMACHC activity. We created stable mammalian cell lines expressing wildtype (WT) and mutant (R132*) MMACHC protein to check the efficacy of readthrough drugs. WT cDNA clone of MMACHC gene was purchased and mutant clone (R132*_MMACHC) was prepared by site-directed mutagenesis. WT and R132*_MMACHC constructs were subcloned into mammalian expression vector (pcDNA5FRT) along with EGFP sequence. Vectors were transfected into Flp-In CHO cell line. Mutant Flp-In cells were treated with G418 and Amlexanox at different concentrations and time intervals. Protein expression was analysed by western blot. Total protein was extracted from Flp-In cell line carrying WT and mutant clones. Western blot showed good expression in WT cells; however, no expression of GFP-tagged protein was observed in mutant (R132*) cells suggesting no expression of MMACHC with this mutation. On treating R132* Flp-In cells with different concentrations of G418, protein expression at 24 hrs was restored at 15μg/ml with relatively higher band intensities at 100-750μg/ml of G418. However, at 72 hrs, protein expression exponentially increased with G418 concentration. Amlexanox also showed restored protein expression at 50μM to 250μM. The observed readthrough effect of G418 and Amlexanox separately on MMACHC nonsense mutation at position R132* indicates the possibility of restoring the function of truncated R132* MMACHC protein