The BTD Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the Jurkat T lymphocyte cell line, with targeted disruption of the BTD gene. This loss-of-function model enables investigation of biotinidase-dependent biotin recycling and its impact on metabolic pathways. The polyclonal format ensures a diverse gene-edited cell pool, minimizing clonal bias while providing robust ablation of biotinidase function.
Jurkat cells are immortalized human T lymphocytes isolated from an acute T cell leukemia patient, widely used to study T cell receptor signaling, cell-mediated immunity, and leukemogenesis. Their suspension growth and genetic tractability make them a standard platform for CRISPR-based gene editing, while their leukemic origin offers a context for exploring metabolic dysregulation in hematological cancers.
Biotinidase, encoded by BTD, hydrolyzes biocytin to release free biotin, a necessary cofactor for biotin-dependent carboxylases such as acetyl-CoA carboxylase, pyruvate carboxylase, propionyl-CoA carboxylase, and 3-methylcrotonyl-CoA carboxylase. These enzymes drive critical carboxylation reactions in fatty acid synthesis, gluconeogenesis, and amino acid catabolism. BTD activity is modulated by biotin concentration and the transcription factor HNF4A, and interacts with biocytin, biotin, and biotinylated peptides within a pathway that includes holocarboxylase synthetase. CRISPR/Cas9-mediated disruption of BTD impairs biotin recycling, consequently reducing carboxylase activities and perturbing metabolic homeostasis.
In the Jurkat T lymphocyte background, BTD knockout provides a physiologically relevant model for biotinidase deficiency??a disorder marked by neurological deficits, skin rash, and metabolic acidosis??and for studying multiple carboxylase deficiency. The leukemic origin of Jurkat cells additionally permits investigation of how biotin metabolic pathways influence cancer cell metabolism, T cell activation, and immune function. Loss of biotinidase may disrupt the provision of biotin to carboxylases, altering anabolic flux and energy metabolism, thereby affecting proliferation and signaling in T cells.
This polyclonal knockout product supports diverse applications: modeling biotinidase deficiency, examining biotin metabolism in T lymphocytes, studying metabolic reprogramming in leukemia, and screening compounds targeting carboxylase deficiencies. Key assays include biotinidase activity measurement, western blotting, RT-qPCR, carboxylase activity assays, metabolic flux analysis, biotin rescue experiments, and flow cytometry. Such methods enable comprehensive functional characterization of BTD disruption. For additional information or to discuss customization, please contact Ascent Research.
