The HMGB1 Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population of HAP1 cells engineered to disrupt the HMGB1 locus. This polyclonal knockout pool provides a heterogeneous collection of gene-disrupted alleles, enabling loss-of-function studies without the need for single-cell cloning. The cells are supplied as a ready-to-use pool, facilitating rapid integration into functional assays.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, retaining the Philadelphia chromosome and BCR-ABL fusion. Its near-haploid genome simplifies genetic manipulation and phenotypic interpretation, making it a widely adopted model for haploid genetic screens and targeted gene knockout. The line’s leukemic origin provides a relevant background for investigating oncogenic signaling and tumor biology.
HMGB1 encodes a highly conserved nuclear DNA-binding protein that functions as both a non-histone chromosomal scaffold and a damage-associated molecular pattern (DAMP). Upon cellular stress or injury, HMGB1 is released extracellularly, where it engages receptors including RAGE, TLR4, and TLR2. This interaction recruits adaptors such as MyD88 and IRAK, leading to IKK-mediated NF-??B activation and downstream transcription of pro-inflammatory cytokines (e.g., IL-6, TNF-??, IL-8) and matrix metalloproteinases (e.g., MMP-9). HMGB1 activity is tightly regulated by upstream signals like TNF-??, IL-1??, LPS, and oxidative stress, and it forms complexes with partners such as p53 and DNA to modulate transcription and genomic stability.
In the HAP1 model, homozygous disruption of HMGB1 is facilitated by haploid genetics, enabling efficient evaluation of its dual intracellular and extracellular roles. This polyclonal knockout population permits dissection of HMGB1-dependent signaling in a human leukemic backdrop, where aberrant inflammatory and survival pathways are critical. Researchers can interrogate how HMGB1 loss impacts NF-??B-driven cytokine secretion, receptor-mediated crosstalk, and cellular responses to genotoxic or inflammatory challenges, directly linking these phenotypes to chronic myeloid leukemia pathophysiology.
Key applications include studying the DAMP-mediated inflammatory response, validating HMGB1 as a therapeutic target in cancer and autoimmune diseases, and screening for upstream regulators or downstream effectors using assays such as ELISA for extracellular HMGB1, Western blot for intracellular localization, NF-??B luciferase reporters, and co-immunoprecipitation of RAGE/TLR4 complexes. The cells also support migration, viability, and proliferation assays for functional profiling. For further technical details or assistance with assay development, please contact Ascent Research.
