The HMGB2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HT29 human colorectal adenocarcinoma cell line. This product provides a genetically heterogeneous pool of cells carrying targeted disruptions in the HMGB2 gene, enabling loss-of-function studies without clonal selection artifacts. The polyclonal format maintains population-level diversity, making the model suitable for experiments where bulk functional readouts are required and where clonal variation may confound phenotypic analyses.
The parental HT29 cell line was established from a primary colon adenocarcinoma of a 44-year-old female and is a classic model of intestinal epithelial physiology and colorectal cancer. HT29 cells possess mutant TP53 and APC genes, while retaining wild-type KRAS and microsatellite stable (MSS) status. This genetic profile recapitulates a subset of sporadic colorectal tumors, making HT29 an appropriate host for studying tumor suppressor pathways, Wnt signaling, and chemotherapeutic responses in a TP53- and APC-deficient context.
HMGB2 encodes a non-histone chromatin protein that binds preferentially to distorted DNA structures, facilitating DNA bending and regulating chromatin architecture, transcription, and DNA repair. Intracellularly, HMGB2 promotes expression of cell cycle regulators such as CCNA2 and CCNB1, and its activity is governed by upstream factors including TP53, NF-??B, SP1, and E2F1 in response to DNA damage or oxidative stress. When released into the extracellular space following cell stress or necrosis, HMGB2 serves as a damage-associated molecular pattern (DAMP) and activates innate immune signaling through RAGE and TLR4 receptors. Downstream of these receptors, MyD88-dependent recruitment of the IKK complex leads to I??B?? phosphorylation and NF-??B nuclear translocation, culminating in the transcriptional induction of pro-inflammatory cytokines (IL-6, TNF) and immune modulators.
In the HT29 background, which harbors TP53 and APC mutations, HMGB2 knockout disrupts the interplay between DNA damage responses, cellular senescence, and inflammatory signaling. This model enables investigation of HMGB2-dependent effects on proliferation, apoptosis resistance, and the senescence-associated secretory phenotype (SASP) in a setting that mirrors key genetic lesions of colorectal tumors, facilitating studies on how HMGB2 modulates chromatin structure and cytokine secretion in a p53-deficient, APC-mutant environment.
Typical applications include western blotting and RT-qPCR for expression analysis, cell viability (MTT/XTT) and apoptosis (Annexin V) assays, migration and invasion (Transwell) assays, and NF-??B luciferase reporter assays. Further downstream approaches comprise ChIP-qPCR for chromatin binding, RNA-seq transcriptome profiling, flow cytometry-based cell cycle analysis, and senescence-associated ??-galactosidase staining, enabling detailed mechanistic studies and drug screening for HMGB2 inhibitors. For further information, please contact Ascent Research.
