The BHMT2 Knockout HCT 116 Polyclonal Cells are a genetically disrupted human cell population generated by CRISPR/Cas9-mediated targeting of the BHMT2 gene. This polyclonal knockout model provides a heterogeneous pool of cells with loss-of-function mutations in BHMT2, enabling functional studies of BHMT2-dependent processes in a colorectal carcinoma background. The product is supplied as a population of polyclonal knockout cells, suitable for bulk analysis without clonal isolation artifacts.
The host cell line, HCT 116, is a widely used epithelial colorectal carcinoma model derived from a male patient. This cell line harbors oncogenic mutations in KRAS (G13D) and PIK3CA (H1047R) and is deficient in mismatch repair due to loss of MLH1 expression. These genetic features render HCT 116 cells relevant for investigating colorectal cancer biology, including genomic instability, signaling pathway dysregulation, and metabolic adaptations.
BHMT2 encodes betaine-homocysteine S-methyltransferase 2, a zinc-dependent enzyme that catalyzes the remethylation of homocysteine to methionine using S-methylmethionine as the methyl donor, yielding S-methylhomocysteine as a byproduct. This reaction is a key node in the methionine cycle and one-carbon metabolism, directly impacting cellular S-adenosylmethionine (SAM) levels and methylation potential. BHMT2 activity is regulated upstream by SAM, nutritional methionine/folate status, and transcription factors such as HNF4A. Downstream, it influences methionine availability, SAM synthesis, DNA methylation patterns, and polyamine biosynthesis. BHMT2 interacts with homocysteine, S-methylmethionine, and the related enzyme BHMT within the methionine salvage network.
In HCT 116 cells, disruption of BHMT2 is particularly relevant for dissecting cancer-associated methionine metabolism and homocysteine homeostasis. Colorectal cancers often exhibit methionine dependency and altered one-carbon metabolism, impacting proliferation, epigenetic regulation, and chemosensitivity. The mismatch repair-deficient background of HCT 116 further models the interplay between genomic instability and metabolic stress. This knockout model enables the study of BHMT2??s role in maintaining methionine pools and SAM-driven methylation, which may influence gene expression and drug responses, including sensitivity to anti-folate agents like methotrexate or 5-fluorouracil.
Researchers can employ this polyclonal BHMT2 knockout population in various experimental contexts. Typical applications include assessing homocysteine and methionine levels via LC-MS/MS metabolomics, evaluating global DNA methylation changes through LINE-1 bisulfite PCR, and conducting cell proliferation assays under methionine restriction. The model is also suitable for drug sensitivity screens with methotrexate or 5-fluorouracil, and for knockout validation by western blotting and RT-qPCR. These studies advance understanding of BHMT2 in cancer metabolism and may inform therapeutic strategies targeting one-carbon metabolism. For further information, please contact Ascent Research.
