The CBS Knockout Huh-7 Polyclonal Cells are a polyclonal population of Huh-7 cells genetically engineered using CRISPR/Cas9 to disrupt the endogenous CBS locus, creating a loss-of-function model for cystathionine beta-synthase. This polyclonal knockout format provides a heterogeneous pool of cells carrying diverse target-gene disruptions, enabling functional studies without the biases of clonal selection. The product is designed for researchers investigating the transsulfuration pathway, hydrogen sulfide signaling, and homocysteine metabolism in a hepatocellular carcinoma background.
The parental Huh-7 cell line is a well-differentiated hepatocellular carcinoma line originally derived from a 57-year-old Japanese male. These adherent epithelial cells maintain key liver-specific functions and serve as a widely used model for hepatocyte biology, drug metabolism, and liver cancer research. Huh-7 cells are particularly relevant for studying hepatic pathways, given their retention of metabolic enzymes and signaling networks characteristic of the liver.
CBS encodes cystathionine beta-synthase, the committing enzyme of the transsulfuration pathway that irreversibly converts homocysteine and serine to cystathionine using a heme cofactor and S-adenosylmethionine as allosteric activator. This reaction is critical for cysteine and glutathione synthesis, as well as for hydrogen sulfide (H?S) production via downstream enzymes such as cystathionine gamma-lyase. CBS expression is transcriptionally regulated by SP1 and NF-Y, with homocysteine and S-adenosylmethionine acting as key metabolic effectors. Loss of CBS function disrupts the conversion of homocysteine to cystathionine, leading to accumulation of homocysteine and depletion of cysteine and glutathione, thus linking this gene to hyperhomocysteinemia, oxidative stress, and vascular pathology.
In Huh-7 hepatocellular carcinoma cells, disruption of CBS provides a physiologically relevant model to dissect the role of transsulfuration and H?S signaling in liver cancer. Hepatocytes are central to methionine and homocysteine metabolism, and alterations in CBS activity have been implicated in liver tumorigenesis, cellular proliferation, and redox homeostasis. The polyclonal CBS knockout Huh-7 cells allow investigation of how loss of CBS function affects hepatoma cell viability, response to homocysteine challenge, and endogenous H?S production, contributing to a better understanding of cysteine auxotrophy and metabolic vulnerabilities in cancer.
These polyclonal knockout cells are suitable for a broad range of experimental applications, including quantification of homocysteine and cystathionine levels via LC-MS, measurement of CBS enzyme activity and H?S production, and assessment of glutathione content and oxidative stress markers. They can be employed to study homocystinuria pathophysiology, evaluate drug candidates for hyperhomocysteinemia, and explore the interplay between one-carbon metabolism and hepatocellular carcinoma. Researchers may also use this model to investigate the impact of CBS loss on cellular responses to chemotherapeutic agents and metabolic inhibitors. For additional details or technical support, please contact Ascent Research.
