The KMT2E Knockout HEK293T Polyclonal Cells product provides a heterogeneous pool of CRISPR/Cas9-edited cells in which the KMT2E gene has been disrupted, generating a functional loss-of-function model for studying the roles of this histone methyltransferase. The polyclonal format encompasses a diverse spectrum of editing events across the cell population, enabling robust assessment of gene disruption phenotypes without clonal selection bias. This product is particularly suited for experiments that require high-throughput functional screening or studies where polyclonal representation reduces the influence of clonal heterogeneity, making it a versatile tool for probing KMT2E-dependent mechanisms in a widely adopted cellular background.
HEK293T is a human embryonic kidney epithelial cell line engineered to stably express the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication. This characteristic confers exceptionally high transfection efficiency, making HEK293T a preferred host for protein overexpression, lentivirus production, and transient reporter assays. Derived from the original HEK293 line, these cells retain an adherent growth morphology and are extensively characterized for their biochemical tractability and rapid proliferation. Their epithelial origin and robust protein expression machinery provide a physiologically relevant context for examining chromatin-modifying enzymes and nuclear signaling events.
KMT2E encodes a histone-lysine N-methyltransferase that catalyzes the monomethylation and dimethylation of histone H3 at lysine 4 (H3K4me1/me2), a chromatin mark associated with active and poised transcriptional states. It functions within a conserved COMPASS-like complex comprising core subunits ASH2L, RBBP5, WDR5, and DPY30, and its catalytic activity is modulated by interactions with HCFC1, OGT, USP7, and MEN1. KMT2E transcriptionally regulates key genes including the HOXA cluster (HOXA9, HOXA10), cell cycle inhibitors (CDKN1A), and apoptotic regulators (BCL2 family members, BIRC5), while receiving upstream inputs from WNT3A, p53-activating stimuli, retinoic acid, NOTCH1, and E2F transcription factors. Through these networks, KMT2E coordinates chromatin accessibility with cellular proliferation, differentiation, and stress responses, often acting in a context-dependent manner as a tumor suppressor or oncogene.
In the HEK293T background, which retains wild-type TP53 and active Wnt/??-catenin signaling, KMT2E disruption allows researchers to dissect its involvement in transcriptional programs governed by these pivotal pathways. The loss of KMT2E-mediated H3K4 methylation can alter the expression landscape of CTNNB1 and TP53 target genes, affecting cell cycle progression and apoptotic thresholds. This cellular backdrop is especially advantageous for examining how KMT2E integrates developmental and stress signals, given the cell line??s responsiveness to transfection-based pathway reconstitution and small-molecule perturbations. As HEK293T is non-tumorigenic yet highly proliferative, it offers a controlled environment to study KMT2E??s role in maintaining genomic stability and epigenetic regulation without the compounding effects of cancer-associated mutations.
This polyclonal knockout model is ideally suited for diverse assays including RNA-seq-based transcriptome profiling to identify KMT2E-dependent gene networks, ChIP-qPCR for mapping H3K4me2 changes at specific promoters, and co-immunoprecipitation to validate protein interactions with components like CTNNB1 or TP53. Functional analyses can be expanded to MTT/BrdU proliferation assays, Annexin V/PI apoptosis detection, and flow cytometric cell cycle analysis to quantify phenotypic outcomes. Moreover, the cells are compatible with dual-luciferase reporter systems for interrogating Wnt and p53 pathway activities, as well as epigenetic compound screening to uncover KMT2E-modulating agents. Sanger sequencing upstream of functional experiments confirms editing efficiency. For further technical details and custom validation requests, please contact Ascent Research.
