The DMGDH Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the DMGDH gene has been disrupted to create a loss-of-function model for dimethylglycine dehydrogenase. This polyclonal product offers a heterogeneous knockout background suitable for population-level analyses of metabolic pathway perturbations without clonal biases.
The host cell line HAP1 is a near-haploid human chronic myeloid leukemia cell line derived from KBM-7 cells, exhibiting fibroblast-like morphology. Its haploid karyotype makes it an ideal platform for genetic perturbation studies, as single-gene disruptions can lead to unmasked phenotypic consequences, simplifying functional genetic analyses.
DMGDH encodes a mitochondrial flavoprotein that catalyzes the oxidative demethylation of dimethylglycine to sarcosine, donating a one-carbon unit to tetrahydrofolate (THF) to form 5,10-methylene-THF. This reaction is pivotal in choline degradation and one-carbon metabolism, linking the betaine-homocysteine methyltransferase (BHMT) pathway and sarcosine dehydrogenase (SARDH)-mediated sarcosine catabolism to the folate cycle and methionine regeneration. DMGDH activity is dependent on FAD cofactor and mitochondrial import machinery, and sits upstream of SARDH in the sequential demethylation of choline-derived methylamines. Its function influences the production of sarcosine, glycine, and one-carbon units required for nucleotide biosynthesis and methylation reactions, including the generation of S-adenosylmethionine (SAM). Key pathway components include choline dehydrogenase (CHDH), betaine aldehyde dehydrogenase (ALDH7A1), BHMT, DMGDH, SARDH, methionine synthase (MTR), and methylenetetrahydrofolate reductase (MTHFR).
In the HAP1 haploid context, disruption of DMGDH eliminates the primary route for dimethylglycine clearance, leading to its accumulation and reduced sarcosine synthesis. This model captures a critical node in one-carbon metabolism and unmasked the phenotypes associated with dimethylglycine dehydrogenase deficiency, enabling direct investigation of metabolic flux and compensatory pathways without interference from a second functional allele.
This polyclonal knockout cell population is suited for a range of research applications, including studies of one-carbon metabolism, choline degradation, methylation capacity, and folate cycle dynamics. Researchers can employ quantitative assays such as dimethylglycine and sarcosine quantification, SAM/SAH ratio measurements, and metabolic flux analysis with stable isotopes to dissect pathway perturbations. Additional applications extend to folate cycle intermediate profiling, enzyme activity assays, and choline deprivation viability experiments. These cells provide a robust model for modeling inborn errors of one-carbon metabolism and screening for modulators of the dimethylglycine dehydrogenase pathway. For more detailed information or technical support, please contact Ascent Research.
