The HMGN3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the HMGN3 gene has been disrupted via CRISPR/Cas9-mediated gene targeting. This heterogeneous pool of edited cells provides a robust loss-of-function model for investigating HMGN3-dependent biological processes without the need for single-cell clone isolation. The polyclonal format captures the diversity of editing outcomes while maintaining functional knockout at the population level, making it particularly suitable for high-throughput screening and assays requiring immediate availability.
The HAP1 host cell line is a near-haploid, fibroblast-like line derived from KBM-7 chronic myeloid leukemia cells. Its adherent growth and near-haploid genome simplify gene knockout studies, as the presence of a single allele reduces the likelihood of compensatory mutations and facilitates efficient CRISPR/Cas9 editing. This genetic simplicity enables clear genotype?Cphenotype correlations and is highly valued in functional genomics research.
HMGN3 encodes a nucleosome-binding protein that reduces chromatin compaction to facilitate transcriptional activation. It interacts directly with histones and nucleosomes, associates with the SWI/SNF chromatin remodeling complex, and is regulated by glucose, cAMP signaling, and the transcription factor PDX1. HMGN3 promotes the expression of insulin, AXIN2, and MYC, thereby linking metabolic and growth signals to gene regulation. In the WNT/??-catenin pathway, HMGN3 interacts with ??-catenin and modulates TCF/LEF target gene expression; upon WNT ligand binding, Frizzled receptor activation leads to DVL-mediated inhibition of GSK3??, stabilization of ??-catenin, and nuclear translocation, where HMGN3 assists in chromatin decompaction at target loci. In pancreatic ??-cells, HMGN3 couples glucose sensing to insulin secretion by modulating chromatin accessibility at the insulin gene.
In the HAP1 near-haploid background, knockout of HMGN3 eliminates potential confounding effects from a second allele, yielding a cleaner loss-of-function phenotype. This model is particularly valuable for dissecting HMGN3??s role in chromatin organization and transcription, as HAP1 cells retain fundamental chromatin regulatory mechanisms. Disruption of HMGN3 is expected to reduce chromatin accessibility at its target genes, impairing transcriptional responses to upstream stimuli such as glucose and ??-catenin signaling. Consequently, the model enables researchers to mechanistically link HMGN3-dependent chromatin changes to altered gene expression programs.
Researchers can employ these knockout cells in a wide range of functional assays. Chromatin accessibility and epigenetic profiling can be performed using ATAC-seq or ChIP-qPCR, while transcriptomic consequences are assessed via RNA-seq and RT-qPCR. Protein-level validation is achieved by western blot and immunofluorescence, and pathway activity can be monitored through reporter assays for WNT/??-catenin or insulin promoter activity. These cells are suited for investigating pancreatic ??-cell biology, type 2 diabetes mechanisms, and WNT signaling dynamics, as well as for drug target validation and screening campaigns. For further information or technical support, please contact Ascent Research.
