The ASH1L Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human ASH1L gene in the near-haploid HAP1 cell line. This loss-of-function model enables the study of ASH1L-dependent chromatin regulation and transcription without the complexity of diploid gene dosage, as the near-haploid background simplifies functional genomics analyses. The polyclonal pool represents a heterogeneous population of edited cells, each carrying distinct gene disruptions introduced by CRISPR/Cas9-mediated gene targeting, providing a powerful tool for pooled functional screening and bulk assays.
HAP1 is an adherent, fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia (CML) clone. Its near-haploid karyotype, stable growth characteristics, and well-characterized origin make it a widely used model in cancer biology and genetic screening. The parental line retains key signaling pathways relevant to hematopoietic malignancies, and its origin from a Philadelphia chromosome-negative CML patient provides a context for studying alternative leukemogenic mechanisms. This background is particularly relevant for studying genes like ASH1L, which has been implicated in acute myeloid leukemia and other hematological disorders.
ASH1L is a histone-lysine N-methyltransferase that catalyzes mono- and dimethylation of histone H3 at lysine 36 (H3K36me1/me2), predominantly at promoter regions. It functions as a transcriptional coactivator downstream of Notch and Wnt pathways: NICD activates ASH1L expression, and ASH1L deposits H3K36me marks to facilitate transcription of HOXA and HOXB clusters. ASH1L interacts with COMPASS-like complex subunits including MLL1, menin, WDR5, RbBP5, ASH2L, and DPY30. Dysregulation of ASH1L-mediated methylation is associated with developmental disorders and leukemic transformation, underscoring its critical role in gene expression.
In the HAP1 leukemia-derived background, ASH1L knockout disrupts the normal histone methylation landscape at key developmental and oncogenic loci. Since HAP1 cells retain functional Notch and Wnt signaling components, this model allows researchers to interrogate how loss of ASH1L alters the transcriptional output of these pathways. Given the near-haploid state, the knockout yields unambiguous loss-of-function phenotypes, making it ideal for dissecting ASH1L-dependent mechanisms in proliferation, differentiation, and drug response. The model is particularly suited for studying the role of ASH1L in acute myeloid leukemia and its intersection with HOX gene dysregulation.
This polyclonal knockout product is optimized for a range of assays including western blotting and RT-qPCR for expression analysis, ChIP-qPCR and RNA-seq for epigenomic and transcriptomic profiling, and histone methylation analysis to directly measure H3K36me changes. It is also applicable for proliferation assays, drug sensitivity screening, and immunofluorescence studies. High-throughput genetic screens using this polyclonal population can uncover synthetic lethal interactions or identify ASH1L-dependent signaling dependencies. For further technical specifications, experimental protocols, or support, please contact Ascent Research.
