The KMT2A Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the KMT2A gene in the human near-haploid HAP1 cell line. This product provides a versatile loss-of-function model for studying the biology of KMT2A, a histone methyltransferase frequently altered in acute leukemias. The polyclonal format consists of a heterogeneous pool of cells harboring CRISPR-mediated gene disruptions, enabling robust pooled analyses while avoiding clonal artifacts. As a research tool, it allows investigations into the epigenetic and transcriptional consequences of KMT2A deficiency without requiring single-cell clone isolation.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and possess a near-haploid karyotype, with most chromosomes present in a single copy. This unique genetic background simplifies functional genomics studies because a single mutagenic event can unmask loss-of-function phenotypes, bypassing the need for bi-allelic inactivation. HAP1 cells retain the BCR-ABL translocation characteristic of chronic myeloid leukemia, providing a disease-relevant context for exploring the interplay between oncogenic kinase signaling and chromatin regulation. Their rapid growth and ease of manipulation make them an ideal host for generating knockout pools.
KMT2A encodes a histone H3 lysine 4 (H3K4) methyltransferase that functions as the catalytic subunit of the COMPASS-like complex, interacting closely with MEN1, ASH2L, RBBP5, and WDR5. It catalyzes H3K4 trimethylation at promoter regions of developmentally critical genes, including the HOXA cluster, MEIS1, and PBX3, thereby maintaining active transcription. KMT2A activity is regulated by upstream signals such as Wnt/??-catenin and CREB, and its methyltransferase function is essential for transcriptional elongation and normal hematopoiesis. Disruption of KMT2A leads to loss of H3K4me3 marks and aberrant silencing of target genes, which can impair cellular proliferation and differentiation programs.
In the HAP1 background, KMT2A knockout is anticipated to cause widespread changes in histone methylation patterns and gene expression, potentially phenocopying aspects of MLL loss-of-function observed in developmental disorders and leukemia. The haploid nature of the cell line ensures that the knockout effect is not masked by a wild-type allele, yielding a clear loss-of-function signature. Moreover, the BCR-ABL-positive status allows researchers to explore crosstalk between dysregulated kinase pathways and epigenetic modifiers, which is particularly relevant for understanding resistance mechanisms in myeloid malignancies.
Researchers can employ this knockout model in a variety of assays, including ChIP-qPCR for H3K4me3 profiling, RNA-seq for transcriptomic analysis, proliferation and differentiation assays, and western blotting for histone marks. It is well-suited for functional genomics screens, epigenetic mechanistic studies, and preclinical drug target validation??especially for agents targeting DOT1L in MLL-rearranged leukemias. The polyclonal population supports high-throughput screening applications and pooled library screens. For further information or ordering, please contact Ascent Research.
