The BCKDK Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from Jurkat T lymphocytes, with targeted disruption of the BCKDK gene to eliminate its kinase activity. This loss-of-function model enables interrogation of BCKDK biological roles in a human T cell context without clonal bias, offering a heterogeneous pool suitable for bulk population studies.
Jurkat cells are an immortalized human T lymphocyte line established from a 14-year-old male with acute T cell leukemia. Widely used to study T cell receptor signaling, apoptosis, and HIV infection, they also model T cell leukemia biology, particularly metabolic adaptation and oncogenic signaling. As a suspension line, they are amenable to high-throughput screening, biochemical assays, and functional genomics approaches.
BCKDK encodes a mitochondrial kinase that inactivates the branched-chain ??-ketoacid dehydrogenase (BCKDH) complex by phosphorylating its E1?? subunit (BCKDHA), impairing the catabolism of branched-chain amino acids (BCAAs) such as leucine, isoleucine, and valine. Elevated BCAAs enhance mTORC1 signaling, a central growth regulator. BCKDK is antagonized by PPM1K phosphatase and transcriptionally regulated by KLF15; insulin and leucine also modulate its activity. Active mTORC1 phosphorylates S6K1 and 4E-BP1 to drive protein synthesis and proliferation.
In Jurkat T lymphocytes, BCKDK knockout disrupts BCKDH inactivation, likely leading to constitutive BCAA catabolism and reduced mTORC1 activity, which can impact T cell activation, proliferation, and survival. Given the leukemic origin of Jurkat cells, this model is valuable for studying metabolic reprogramming in T cell leukemia, where mTORC1 is often hyperactive. Additionally, since BCKDK mutations are linked to autism spectrum disorder and intellectual disability, this knockout provides a cell-based platform for neurodevelopmental research.
Researchers can employ this knockout population to investigate BCKDK-dependent pathways via Western blotting for BCKDK and phospho-BCKDHA, BCAA quantification, and phospho-S6K1 measurement for mTORC1 activity. Functional assays include MTT proliferation, flow cytometry for T cell markers, and Annexin V apoptosis staining. This model also supports BCKDK inhibitor screening for cancer and metabolic disorder therapeutics. For further technical details or custom applications, please contact Ascent Research.
