This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KMT2B gene in Jurkat cells. The polyclonal format preserves the genetic heterogeneity inherent to a mixed population of edited cells, avoiding clonal selection while achieving gene disruption at the population level. KMT2B encodes a histone-lysine N-methyltransferase 2B that installs mono- and trimethylation on histone H3 at lysine 4 (H3K4me1/me3), a chromatin mark tightly associated with active transcriptional states. Disruption of this epigenetic writer in a T-lymphoblast background offers a physiologically relevant loss-of-function model for investigating chromatin-dependent gene regulation, T-cell biology, and leukemogenic processes driven by aberrant histone methylation.
The Jurkat host cell line is an extensively characterized human T-cell leukemia model derived from the E6.1 clone. These cells grow in suspension and retain key features of T-lymphoblasts, including T-cell receptor (TCR) signaling competence, immune surveillance functions, and the capacity for cytokine production upon activation. Jurkat cells are widely used to study TCR-proximal signal transduction, NFAT and NF-??B activation, and the molecular basis of T-cell acute lymphoblastic leukemia (T-ALL). Their well-documented genetic and signaling landscape makes them an ideal recipient for disruption of chromatin modifiers like KMT2B, where the downstream consequences on gene expression programs can be systematically interrogated in a cell-autonomous manner.
KMT2B functions as the catalytic subunit within the multiprotein COMPASS (Complex of Proteins Associated with Set1)-family methyltransferase complexes, where it partners with core subunits such as ASH2L, WDR5, RBBP5, and DPY30, and is regulated by menin and other MLL complex components. Upstream, KMT2B expression is modulated by transcription factors TCF3 and TCF4, and by developmental signaling cascades including Notch and Wnt pathways. Once recruited to chromatin, KMT2B catalyzes H3K4me3 deposition at promoter regions, which in turn recruits transcriptional coactivators and promotes expression of critical downstream targets, most notably clustered HOX genes and cell cycle regulators. The methyltransferase thus acts as a central node linking extracellular signals, chromatin modification, and transcriptional output, with direct implications for cell identity and proliferation.
In the Jurkat T-cell context, KMT2B knockout disrupts the normal epigenetic landscape governing immune response genes. H3K4me3 profiles at enhancers and promoters of TCR signaling components, cytokine genes, and transcription factors are expected to shift, potentially altering T-cell activation kinetics, proliferation, and cytokine production. This model is particularly relevant for dissecting the role of MLL complex dysfunction in T-ALL, where rearrangements or deregulation of KMT2 family members contribute to leukemogenesis. Furthermore, because KMT2B mutations are linked to Kabuki syndrome and other developmental anomalies, the knockout cells enable structure?Cfunction studies that connect specific methyltransferase activities to pathological gene expression programs.
Researchers can employ this knockout polyclonal population in a broad spectrum of assays to probe epigenetic regulation in T cells. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) for H3K4me3 permits genome-wide mapping of methylation changes, while RNA-seq delivers quantitative transcriptomic profiles that reveal differentially expressed genes, including those of the HOX clusters. Complementary techniques such as qPCR, Western blot analysis of histone modifications, and flow-cytometric detection of T-cell activation markers (e.g., CD69, CD25) provide robust validation readouts. Proliferation and viability assays further assess functional consequences of KMT2B loss. These applications make the product a powerful tool for academic and pharmaceutical research into epigenetic therapies and leukemia biology. For additional information, please contact Ascent Research.
