The KMT2A Knockout A-549 Polyclonal Cells product consists of a population of CRISPR/Cas9-edited polyclonal knockout cells derived from the human A-549 lung adenocarcinoma epithelial cell line. The cells carry a CRISPR/Cas9-mediated disruption of KMT2A, abolishing its histone methyltransferase function. This polyclonal pool represents a diverse collection of gene-edited cells, enabling robust loss-of-function studies without reliance on single clonal isolates. The knockout model is suitable for investigating the role of KMT2A in transcriptional regulation and cancer biology.
The A-549 cell line is a widely utilized model of human lung adenocarcinoma, originally isolated from a 58-year-old Caucasian male. A-549 cells exhibit epithelial morphology and are characterized by the expression of lung adenocarcinoma markers, making them a standard system for studying lung cancer biology, drug responses, and oncogenic signaling pathways. Their robust growth and ease of manipulation render them ideal for CRISPR-based genetic modification, allowing researchers to dissect gene function in a disease-relevant context.
KMT2A encodes the catalytic subunit of the MLL histone methyltransferase complex, which includes core components MEN1, ASH2L, RBBP5, WDR5, and DPY30, along with interacting factors such as PSIP1/LEDGF, CHD8, HCFC1, CREBBP, EP300, and DOT1L. This complex catalyzes mono-, di-, and trimethylation of histone H3 at lysine 4 (H3K4me1/2/3) at the promoters of target genes, most notably the HOXA and HOXB gene clusters. KMT2A function is positively regulated by upstream signals including WNT ligands through ??-catenin/TCF, NOTCH1 intracellular domain, retinoic acid receptors, and the kinase CDK9. The methyltransferase activity of KMT2A directly promotes the expression of downstream targets such as HOXA genes, MYC, and BCL2, and represses cell cycle inhibitors CDKN1A and CDKN2B. Through these interactions, KMT2A orchestrates key programs in development, hematopoiesis, and oncogenesis.
In A-549 lung adenocarcinoma cells, disruption of KMT2A abolishes MLL complex-mediated H3K4 methylation at critical genomic loci, thereby impairing transcriptional activation of pro-proliferative and anti-apoptotic pathways. This loss-of-function model enables researchers to dissect the contribution of KMT2A-dependent epigenetic regulation to the malignant phenotype of lung adenocarcinoma. Given the involvement of KMT2A in hematopoietic malignancies such as acute myeloid leukemia and acute lymphoblastic leukemia, and its emerging roles in solid tumors, the knockout cell model bridges studies of chromatin remodeling and lung cancer biology.
Researchers can employ this knockout cell population in a diverse array of assays, including Western blotting to assess global H3K4 methylation levels, RT-qPCR to quantify expression changes in HOXA target genes, and RNA-seq for whole-transcriptome profiling. Chromatin immunoprecipitation followed by qPCR (ChIP-qPCR) allows verification of KMT2A occupancy at target promoters, while functional assays such as proliferation, apoptosis, and flow cytometric cell cycle analysis enable characterization of phenotypic consequences. The polyclonal knockout cells are also suitable for drug target validation studies and high-content screening. For further details, please contact Ascent Research.
